The ultrastructure and development of tubular and crystalline P-protein in the sieve elements of certain papilionaceous legumes

Summary The ultrastructure of the primary sieve elements of several papilionaceous legumes was studied using hypocotyl and young internode segments fixed in glutaraldehyde followed by osmium tetroxide. In particular, the study sought to determine whether the crystalline flagellar inclusions characteristic of these species are developmentally related to the P-protein bodies present in the phloem of these and other legumes and of angiosperms generally. The crystalline inclusions consist of a central body terminated at one or both ends by a gradually tapering tail. The central body is usually spindle shaped in longitudinal section and square in cross section. In all species examined, the inclusion is first seen as a small, thin crystal in the cytoplasm of young sieve elements. The crystal enlarges and acquires tails as the sieve element develops. In certain species, exemplified byDesmodium canadense, numerous tubules are formed in the cytoplasm near the crystal and appear to be concerned in its growth. The observations on the structure and interactions of these two components, tubules and crystalline inclusions, suggest that both represent forms of P-protein: the tubules are continuous with the crystal and are striated like the crystal near the tubule-crystal junction, suggesting that they are adding onto the crystal body; the tubules closely resemble the P-protein tubules described in the literature in that they measure 157 Å in diameter, accumulate in spindle-shaped bundles, and disperse into striated fibrils late in the ontogeny of the sieve element; and finally, the crystal also disperses into fine filaments. The crystalline inclusion therefore probably represents still another aggregation state of P-protein, one that is characteristic of papilionaceous legumes. The different stages of crystal aggregation and the diverse forms of P-protein now known are discussed briefly in relation to the control of macromolecular assembly and subunit packing.     

Ultrastructure of P-protein inHevea brasiliensis during sieve-tube development and after wounding

Summary P-protein and the changes it undergoes after wounding of sieve tubes of secondary phloem in one- to two-year old shoots ofHevea brasiliensis has been studied using electron microscopy. The P-protein in the form of tubules with a diameter of 8–9 nm and a lumen of 2–2.5 nm occurred in differentiating sieve elements and appeared as compact bodies which consisted of small aggregates of the tubules. As the sieve elements matured, these P-protein bodies dispersed with a disaggregation of the tubules before they turned into striated fibrils, 10–11 nm in diameter. In wounding experiments, as the mature sieve elements collapsed after cutting, their striated P-protein converted into tubules. These tubules were the same in ultrastructure as the tubules in differentiating sieve elements and they often were arranged in crystalline aggregates.     

Structure and significance of cruciate flagellar root systems in green algae: Zoospores ofUlva lactuca (Ulvales,Chlorophyceae)

The ultrastructure of the flagellar apparatus in the quadriflagellate zoospores ofUlva lactuca was examined. The two L-shaped pairs of basal bodies are arranged in mirror image relation. Two apical capping plates connect adjacent basal bodies of different pairs with each other. The flagellar root system is cruciate and exhibits a microtubular part (4-2-4-2 system) and a complex and elaborate fibrillar part. The latter consists of two striated fibres (striation pattern 32 nm) closely associated with the two-stranded roots and four differently patterned fibres (striation pattern 150–160 nm) which are more internally located and run parallel to all four microtubular roots. The presence of four microtubular roots and six striated fibres is at present not known for any other green alga and taxonomic implications are discussed.     

Development of P-protein in sieve elements ofMimosa pudica

Summary The P-protein in sieve elements of leaves ofMimosa pudica L. is first discernible as fine fibrous material which forms homogeneous aggregates. Ribosomes, rough endoplasmic reticulum, and dictyosomes with associated vesicles occur in the cytoplasm surrounding the aggregates. The plastids and mitochondria are in a parietal position in the parts of the cell where the nascent P-protein accumulates. In a later stage, the fibrillar material is organized into a three-dimensional system of five- and six-sided elongated compartments. The corners of the compartments appear solid at first, then they become electron lucent in the center and assume tubular form. Aggregates of mature P-protein tubules usually occur near the compartmentalized system. Tubules in pentagonal or hexagonal arrangements may be present in the aggregates and may be partly interconnected. The conclusion was drawn that the P-protein tubules are assembled at the corners of compartments within a continuous orderly system. The fully formed tubules occur first in aggregates, the P-protein bodies. Later the aggregates become loose and partly dispersed. Many of the dispersed tubules assume a loose, extended, helical form characteristic of P-protein in older sieve elements.This work was supported in part by National Science Foundation grant GB-5506. I am also grateful to MissHatsume Kosakai and Mr.Robert H.Gill for technical assistance.     

Foliar anatomy ofPolygonum (Polygonaceae): Survey of epidermal and selected internal structures

Several anatomical characters in leaves were described, and their distribution determined, for 153 species ofPolygonum, mostly from herbarium specimens. Structures surveyed were epidermal (glandular and nonglandular trichomes, nodules, specialized parenchyma, stomatal apparatus) and internal (cavities, crystals, laticifer-like cells, nodules, subepidermal fibers). Cleared leaves were supplemented by resin-embedded sections and SEM preparations of selected species. No feature defines any taxonomic section, but some features occur only within one section. Laticifer-like cells, epidermal and internal nodules, resin cups, and unique epidermal and subepidermal cavities seem to be unknown elsewhere; other features (invaginated epidermal cells; enlarged crystal cells confined to paraveinal layer) are rare among angiosperms.     

Karyotype and C-banding inTrigonella sectionFoenumgraecum (Fabaceae)

The six species of the sectionFoenum-graecum ofTrigonella have the same chromosome number, 2n = 16.T. gladiata andT. cariensis have fairly symmetrical karyotypes, while those ofT. foenum-graecum, T. berythea, T. macrorrhyncha andT. cassia are asymmetrical. C-bands are present in all six species but the number of bands and their positive vary considerably among the species. The karyotype evidence suggests that none of the available species of theFoenum-graecum section can be considered as the wild progenitor of fenugreek.     

Monoclonal antibodies against phloem P-protein from plant tissue cultures. II. Taxonomic distribution of cross-reactivity

P-protein, a filamentous protein found in the sieve elements of most angiosperms, is believed to function in the sealing of phloem wound sites. We report here on the use of a highly sensitive immunomicroscopy assay to study the ability of P-protein specific monoclonal antibodies RS21, RS22, and RS23, made against the P-protein from Streptanthus tortuosus (Brassicaceae), to recognize the native P-protein in a number of different plant genera. RS21, RS22, and RS23 all recognized the P-protein in other genera within the Brassicaceae including Arabidopsis and in the closely related family, Capparaceae. RS21 and RS22 also were able to bind to the P-protein in plants more distantly related to S. tortuosus. The labeling of P-protein was also observed in the monocots Iris and Narcissus probed with RS21. No label was seen with members of the Poaceae that are reported to lack P-protein. None of the monoclonal antibodies was able to bind to the P-protein in members of the Cucurbitaceae.     

Distyly and pollen dimorphism inLinum suffruticosum (Linaceae)

The Mediterranean distylousLinum suffruticosum has dimorphic pollen. That from short-styled plants has sub-monomorphic surface excrescences with minute papillae; that from long-styled plants has strongly dimorphic excrescences with conspicuous papillae. The differences are much like those described in species of other sections of the genus. Pollen of the related homostylousL. tenuifolium has monomorphic pollen. It combines characteristics of bothL. suffruticosum types but resembles that of the long-styled plants somewhat more closely. Ribbonlike styles of long-styled plants ofL. suffruticosum provide another feature which seems to be controlled by the S supergene thought to be associated with distylic incompatibility. The pollen ofL. suffruticosum andL. tenuifolium is very similar to that of some species of sectionLinastrum and would support their placement in that section.     

Systematics ofJusticia sect.Pentaloba (Acanthaceae)

The transfer of the four taxa ofSiphonoglossa sect.Pentaloba toJusticia is proposed. It is shown that the taxa of this section were placed inSiphonoglossa primarily because of a single-character phenetic relationship and that they correctly belong inJusticia. In addition to morphology, data from cytology and flavonoid chemistry are also presented that support this intergeneric transfer. A key to the taxa and a detailed taxonomic treatment of the section are provided.     

Electron microscope investigation of sieve-element ontogeny and structure inUlmus americana

Summary During advanced stages of sieve-element differentiation inUlmus americana L., dispersal of the P-protein (slime) bodies results in formation of a peripheral network of strands consisting of aggregates of P-protein components having a striated, fibrillar appearance. The tonoplast is present throughout the period of P-protein body dispersal. Perforation of the sieve plates is initiated during early stages of P-protein body dispersal.Small P-protein bodies consist of tubular components, most of which measure about 180 Å in diameter. With increase in size of the P-protein bodies narrower components appear. At the time of initiation of P-protein body dispersal, most of the components comprising the bodies are of relatively narrow diameters (most 130–140 Å) and have a striated, fibrillar appearance. Both wide and narrow P-protein components are present throughout the period of sieve-element differentiation and in the mature cell as well, and a complete intergradation in size and appearance exists between the two extremes. Both extremes of P-protein component have a similar substructure: an electron-transparent lumen and an electronopaque wall composed of subunits, apparently in helical arrangement. The distribution of P protein in mature sieve elements was quite variable.The parietal layer of cytoplasm in matureUlmus sieve elements consists of plasmalemma, endoplasmic reticulum cisternae in two forms (as a complex network closely applied to the plasmalemma and in stacks along the wall), mitochondria, and plastids.     

Pachyphragma andGagria (Brassicaceae) Revisited: Molecular data indicate close relationship toThlaspi S.Str.

The systematic position ofPachyphragma macrophyllum (Brassicaceae) is unsettled. This species was either treated as a monotypic section ofThlaspi s.l. or was treated as a distinct genus.Gagria lobata apparently closely related toPachyphragma, was also given generic rank. To unravel the phylogenetic relationships ofPachyphragma andGagria, ITS sequence studies were conducted using as a reference representatives of the main lineages ofThlaspi s.l. The molecular data indicate thatGagria lobata andPachyphragma macrophyllum are closely related to members ofThlaspi s.str., an assemblage of species well separated from the otherThlaspi s.l. lineages.     

Expression of the phloem lectin is developmentally linked to vascular differentiation in cucurbits

The conducting elements of phloem in angiosperms are a complex of two cell types, sieve elements and companion cells, that form a single developmental and functional unit. During ontogeny of the sieve element/companion cell complex, specific proteins accumulate forming unique structures within sieve elements. Synthesis of these proteins coincides with vascular development and was studied in Cucurbita seedlings by following accumulation of the phloem lectin (PP2) and its mRNA by RNA blot analysis, enzyme-linked immunosorbent assay, immunocytochemistry and in␣situ hybridization. Genes encoding PP2 were developmentally regulated during vascular differentiation in hypocotyls of Cucurbita maxima Duch. Accumulation of PP2 mRNA and protein paralleled one another during hypocotyl elongation, after which mRNA levels decreased, while the protein appeared to be stable. Both PP2 and its mRNA were initially detected during metaphloem differentiation. However, PP2 mRNA was detected in companion cells of both bundle and extrafascicular phloem, but never in differentiating sieve elements. At later stages of development, PP2 mRNA was most often observed in extrafascicular phloem. In developing stems of Cucurbita moschata L., PP2 was immunolocalized in companion cells but not to filamentous phloem protein (P-protein) bodies that characterize immature sieve elements of bundle phloem. In contrast, PP2 was immunolocalized to persistent ␣ P-protein bodies in sieve elements of the extrafascicular phloem. Immunolocalization of PP2 in mature wound sieve elements was similar to that in bundle phloem. It appears that PP2 is synthesized in companion cells, then transported into differentiated sieve elements where it is a component of P-protein filaments in bundle phloem and persistent P-protein bodies in extrafascicular phloem. This differential accumulation in bundle and extrafascicular elements may result from different functional roles of the two types of phloem. Received: 31 July 1996 / Accepted: 27 August 1996     

3种淡水布纹藻(硅藻纲)的超微结构研究

该研究利用光学和扫描电子显微镜对3种淡水布纹藻(钝布纹藻、库氏布纹藻和赛欧托布纹藻)的形态学进行了研究,以明确它们的精细结构特征;并分析讨论它们在中国的地理分布区域。结果表明：(1) 3种淡水布纹藻皆属于粗糙布纹组;从壳面观看,都具有S形和反S(S)形两种壳瓣外形。(2) 钝布纹藻具有弱的S形壳面外形、T形的外近端壳缝末稍和直的内近端壳缝末稍以及椭圆形的中央区;库氏布纹藻具有明显的S形壳面外形、略微向不同侧偏转的外近端壳缝末稍和直的内近端壳缝末稍以及椭圆形的中央区;赛欧托布纹藻具有弱的S形壳面外形、强烈钩状的外近端壳缝末稍和T形的内近端壳缝末稍以及倾斜的中央区。(3) 钝布纹藻在广东、广西、湖南、四川和云南等地有分布;库氏布纹藻在新疆和湖南等地有发现;赛欧托布纹藻分布于武陵山区,该文是其在中国淡水分布的首次报道。     

EXTERNAL SPORE MORPHOLOGY AND TAXONOMIC AFFINITIES OF PHYLLOGLOSSUM DRUMMONDII KUNZE (LYCOPODIACEAE)

The external spore morphology of Phylloglossum drummondii was studied with the scanning electron microscope and shown to share a number of features with spores in the subgenus Urostachys in Lycopodium. Within Urostachys the affinities of P. drummondii were less evident: the foramino-fossulate distal face, pyramidal proximal surface, and subcircular outline of its spores are similar to spores in the section Phlegmaria while the weakly foveolate proximal surface and raised laesurae are characteristic of spores in the section Selago.     

Sero-systematics ofCytisus sect.Trianthocytisus (Fabaceae)

The serological reaction of seed proteins provides evidence for a partly new systematic arrangement ofCytisus sect.Trianthocytisus and ofCytisus s.l. Proposed modifications agree with recent advances in morphological taxonomy. Sect.Trianthocytisus includes only two species,C. villosus andC. aeolicus. Its position is central within the genus, and this fact agrees with the proposed retypification ofCytisus (type species:C. villosus).C. emeriflorus, formerly included in the same section, constitutes the monospecific sect.Emeroides, which is intermediate towards the genusLembotropis. This is serologically isolated and includes onlyL. nigricans. It is confirmed thatC. sessilifolius should be removed from the genusCytisus as a monospecific genus:Cytisophyllum Lang which is closely allied toHesperolaburnum and toPodocytisus, the most primitive genera ofGenisteae.     

A new species ofJusticia sect.Ansellia (Acanthaceae) from Tanzania

Justicia brevipedunculata, a new species ofJ. sect.Ansellia endemic to Tanzania, is described and illustrated. Detailed palynological information is given, and relationships to other species of the section are discussed.     

Systematic significance of seed coat morphology inSilene L. s. str. (sileneae-caryophyllaceae) from Korea

Seeds of eight species ofSilene s. str. in Korea were examined with a dissecting stereomicroscope and a scanning electron microscope (SEM) to evaluate the systematic significance of seed coat microstructure. The seeds ofSilene s. str. were characterized by triangular or reniform shape, planed or concave dorsal surface in outline, tuberculate or colliculate testa, and sinuated cell margin. Based on the degree of grooves and invagination at dorsal surface, the seeds of the examined taxa ofSilene s. str. fall into two distinct groups: (I) dorsal grooves inconspicuous, surface planed (Silene jenisseensis andS. repens), (II) dorsal grooves conspicuous, surface concave (six remaining taxa). Within each group, seeds of all species may be further distinguished based on shape, size, and testa surface sculpturing. A key to the different species or groups of species is provided, and the systematic significance between/among the taxa is also briefly discussed.     

The musculature of horsehair worm larvae (<Emphasis Type="Italic">Gordius aquaticus</Emphasis>,<Emphasis Type="Italic"> Paragordius varius</Emphasis>, Nematomorpha): F-actin staining and reconstruction by cLSM and TEM

The musculature of larvae of Gordius aquaticus was investigated by laser-scanning microscopy and compared to transmission electron microscopic data for the larva of Paragordius varius. In the anterior portion of the body, the preseptum, four different muscle groups can be distinguished: (1) 12 anterior parietal muscles in the body wall, (2) six oblique muscles that function as retractors of the introvert, (3) six proboscideal muscles, which function as retractors for the proboscis, and (4) six muscles associated with spines of the outermost of the three rings of spines. The posterior portion of the body, the postseptum, possesses four pairs of longitudinal muscle strands in G. aquaticus, the postseptal parietal muscles, that are located dorsolaterally and ventrolaterally. These are not clearly visible in P. varius, where instead three pairs of parietal muscles are present. Additional small muscles are associated with the terminal spines and with the duct running from the pseudointestine to the body wall. All fibers show a cross-striated pattern although this striation is less obvious at the ends of the fibers.     

A revision of Salvia sect. Ekmania (Lamiaceae)

A taxonomic treatment ofSalvia sect.Ekmania, a group of tall shrubs endemic to the Caribbean island of Hispaniola, is provided. Morphology, phenology, pollination biology, habitats, and distributions are discussed. Species boundaries are examined using phenetic analysis of morphological data. Eight species are here accepted as belonging to the section. Two recently described species,S. lavendula andS. paryskii, are considered for inclusion in the section, but only the latter appears to be a member.Salvia lachnaiclada andS. ottoschulzii are treated as conspecific.Salvia bahorucona is recognized as a distinct species.     

Two new species and a new combination in Frantzia (Cucurbitaceae)

The monotypic genusPolakowskia is reduced to a section ofFrantzia. Two new species from Costa Rica,F. villosa andF. talamancensis, are described. The new nomenclatural combination,F. tacaco (Pittier) Wunderlin, is proposed.