Structure and Histochemistry of Stomata and Epidermal Cells in Five Species of Polypodiaceae

The epidermal structure of the five species of ferns, Arthromeriswallichiana (Spr.) Ching., Drymoglossum piloselloides (Prest.),Drynaria quercifolia (L.) J. Smith, Lepisorus nudus (Hook.)Ching. and Pyrrosia nuda (Gies.) Ching., has been investigated.Fifteen types of stomatal structures have been identified ofwhich copolo-desmocytic and coperi-desmocytic are new types.Four more possible stomatal structures: ccpolo-peri-, codesmo-polo-,codesmo-peri- and duplodesmocytic, are suggested. Localizationof starch, insoluble polysaccharides, protein and lipids hasbeen examined histochemically in the guard cells, subsidiarycells and epidermal cells. In Drynaria starch plastids and plastidscontaining both starch and protein are present in guard cells.Starch plastids are present in the subsidiary cells of all speciesexcept in Arthromeris, whereas, they are present in epidermalcells of only Drymoglossum and Lepisorus. Granular or amorphousinsoluble polysaccharides (other than starch) are present inguard cells of all the species, in the subsidiary cells of Arthromeris,Drynaria and Pyrrosia, and in the epidermal cells of Pyrrosia.Except in Pyrrosia lipids are present in the guard cells. Subsidiarycells of Drynaria and the epidermal cells of Arthromeris andDrynaria show lipid bodies. The presence of plasmodesmata andectodesmata is demonstrated in the epidermal cells of Drymoglossum.     

恒河猴皮肤干细胞的体外培养纯化与鉴定

探索恒河猴皮肤干细胞的体外培养及纯化条件,为进一步的研究奠定基础. 通过组织块培养法和消化培养法 在体外培养恒河猴表皮细胞,然后用Ⅳ型胶原吸附法吸附20 min,获得快吸附细胞. 对快吸附细胞进行克隆培养,并进行免疫细胞化学双标染色、RT PCR鉴定 β1 整合素和角蛋白15的表达,用流式细胞仪鉴定纯化前后的细胞中 β1 整合素和角蛋白15的阳性细胞比例,并通过透射电镜观察细胞的超微结构. 组织块培养法和消化培养法均可获得表皮细胞,Ⅳ型胶原纯化后的细胞胞体较小,饱满,核/浆比例大,细胞镶嵌状排列. 细胞克隆分析显示,细胞全克隆生长率高. 细胞免疫荧光显示,分选后的细胞显示 β1 整合素和角蛋白15阳性. RT PCR检查呈现 β1 整合素和角蛋白15的特异性片段. 流式细胞仪检查显示,纯化前的细胞中角蛋白15阳性细胞占总细胞中的比例为8％, β1 整合素阳性细胞的比例为10.7％;纯化后,角蛋白15阳性细胞的比例为89.4％, β1 整合素阳性细胞的比例为88.5％. 通过组织块培养法和消化培养法均可培养获得活性良好的表皮细胞,Ⅳ型胶原吸附法是一种简便、有效的皮肤干细胞分离方法,可以为进一步的眼表上皮替代重建眼表提供足量的高纯度的干细胞建立可靠的物质基础.     

Effects of Acriflavine on the Formation of the Plasma Membrane of Escherichia coli

When cells of acriflavine-sensitive (acrA) and acriflavine-resistant(acrA⁺) Escherichia coli K-12 strains were treated with a ratherhigh concentration (100 µg ml^-1) of acriflavine in mediumthat had been adjusted to pH 8.1, distinct whirlpool-like structuresderived from the plasma membrane appeared not only in the acrAcells but also in the acrA⁺ cells. Chemical analysis was performedto determine the lipid composition of the cells by thin-layerchromatography on silica gel and gas-liquid chromatography.The amount of total fatty acids was significantly higher inthe acrA cells than in the acrA⁺ cells, when cells were culturedin the presence of acriflavine. This difference seems to becaused by the greater accumulation of unsaturated fatty acids(palmitoleic and cis-vaccenic acid) in the acrA mutant cellsthan in the acrA⁺ cells and by the acceleration of this accumulationas a result of the presence of the dye. A comparison of phospholipidcontents between the acrA and acrA⁺ cells cultured under acriflavine-freeconditions showed that the former cells contained more phosphatidylethanolamine(PE) and, in particular, more cardiolipin (CL) than the lattercells. However, the situation was reversed in the case of phosphatidylglycerol(PG). Addition of acriflavine to the medium led to a markedincrease in levels of PE and CL in both acrA and acrA⁺ cellsbut an increase in levels of PG was found only in the acrA⁺cells. (Received October 13, 1992; Accepted May 31, 1993)     

The Effect of Light and Darkness Upon Infection of Asterionella formosa Hassall by the Chytrid Rhizophydium planktonicum Canter emend

Observations, both experimental and microscopic, indicate thatzoospores of Rhizophydium planktonicum Canter emend., can remainalive, but rarely become adherent upon cells of Asterionellaformosa Hassall under conditions of very low light or in completedarkness. The behaviour patterns of zoospores towards host cells underdarkened conditions were compared with those which took placeunder well illuminated conditions leading to the settlementof zoospores on host cells. The differences noted may help toexplain the lack of zoospores found upon inadequately illuminateddiatom cells. Some evidence suggests that young zoospores lack the abilityto adhere to host cells. After the encystment of zoospores uponAsterionella cells in the light, their further growth and developmentcan continue in darkness. Rhizophydium planktonicum Canter emend., Asterionella formosa Hassall, chytrid, diatom, infra-red illumination, zoospore     

A new Dol-P-Man:protein O-D-mannosyltransferase activity from Saccharomyces cerevisiae

The deletion of the protein mannosyltransferase 1 gene (PMT1)of Saccharomyces cerevisiae results in viable cells. O-Mannosylationof proteins is reduced to about half of the value in comparisonto wild-type cells. In order to distinguish between the thePMT1 gene product (= Pmt1p) and residual transferase activity,an in vitro assay to measure Dol-P-Man:protein mannosyltransferaseactivity in cells deleted for PMT1 has been developed. The transferaseactivity of these cells exhibits a pH optimum of 6.5 as comparedto pH 7.5 for Pmt1p. The K_$$$ value of the residual enzyme activityfor the hexapeptide YNPTSV is 7 times higher than that of Pmt1pand shows a clear preference for the seryl/residue. Differencesin substrate affinities as well as in seryl/threonyl preferencebetween the two enzymes, however, depend on the specific sequenceof the peptides used in the enzyme assay. The new enzyme activityshows a significantly lower thermal stability as compared toPmt1p. glycoprotein O-glycosylation mannosyltranferase Saccharomyces cerevisiae     

Biogenic Opal in Three Species of Gramineae

Particulate biogenic opaline silica is concentrated in cellwalls, intercellular deposits and cell lumina of all portionsof the above-ground plant body of three species of PanicoidGramineae,Andropogon gerardi, Sorgastrum nutans and Panicumvirgatum. Morphologically distinct opal phytoliths form notonly in long cells, short cells, trichomes, stomatal elementsand bulliform cells of the epidermis but also within the cellularstructure of mesophyll, vascular, and sclerenchyma tissues.Roots and rhizomes contain measurable quantities of opalinesilica, and phytoliths develop in epidermal long cells, saddle-shapedshort cells, vascular cells, and intercellular deposits. A morphologicallyunique plate-phytolith, formed by silicification of the innertangential wall of the endodermis, is present in the roots ofall three species. Differences in the quantity of opaline silicaoccur between species and between parts of the same species.The amount of opal deposited in the soil annually by root systemsand above-ground parts is approximately equal in magnitud Andropogon gerardi, Sorgastrum nutans, Panicum virgatum, opaline silica deposition     

Infection and Root-Nodule Development in Stylosanthes Species by Rhizobium

Root nodules of the tropical forage legume Stylosanthes occurredonly at lateral root junctions and resulted from direct invasionby rhizobia through spaces between epidermal cells. No infectionthreads were present in either the root hairs or nodules. Invasionof the host cortical cells was through structurally alteredcell walls. The bacteria reached the site of nodule initiationin the lateral root cortex by progressive collapse of the initiallyinvaded cells which were compressed by neighbouring cells toform intercellular thread-like infection zones. The bacteriamultiplied in the invaded cells of the nodule initial whichdivided repeatedly to form the nodule. Bacteroids formed onlywhen the host cells ceased to divide. Some abnormal associations occurred in S. capltata and S. hamata40264A. Division of invaded cells was restricted in S. capitataand the bacteria became enlarged and grossly deformed. In S.hamata restricted cell division was immediastely followed bythe brcakdown of the host cells and, although the bacteria multiplied,no bacteroids were formed. Bacteria isolated from these nodulesformed both effective and abnormal nodules when inoculated ontothe same host.     

A Role for the acrA Gene in the Protection of Escherichia coli Cells against Excess Sodium

The acrA gene determines the sensitivity of Escherichia coliK-12 cells to acriflavine, which is one of the acridine dyesand which effectively eliminates certain plasmids from the bacterialcells. The acriflavine-sensitivity mutation leads to instabilityof plasmids, such as sex (F)- and drug-resistance (R)-factorsand to loss of a membrane protein with molecular weight about60 kDa (Nakamura 1974, 1976, Nakamura et al. 1975, 1981). Wehave found that cells with a mutant acrA gene were also moresensitive to an excess of sodium ions in the medium than werethe wild-type acrA⁺ cells (Nakamura 1977). The product of theacrA gene hindered the accumulation of sodium by the cells.Although mutations in theproA or proB genes, which determinethe synthesis of the enzymes y-glutamyl phosphate reductaseand y-glutamyl kinase, respectively, in the proline-biosyntheticpathway also led to sensitivity to and accumulation of excesssodium, the presence of the acrA⁺ allele decreased both theseparameters. (Received November 1, 1989; Accepted April 27, 1990)     

Electron Transport Coupled to Nitrate Reduction in a Photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans

The electron transport system involved in nitrate reductionand its relationship to photosynthetic cyclic electron transportin a photodenitrifier, Rhodopseudomonas sphaeroides forma sp.denitrificans, were studied. Nitrate oxidized only b-type cytochromein the presence of cyanide, which inhibits nitrite reductase.Heptylhydroxyquinoline-N-oxide (HOQNO) inhibited the oxidationof b-type cytochrome by nitrate, but not the oxidation of b-and c-type cytochrome by nitrite. The inhibition by HOQNO wasovercome by phenazine methosulfate (PMS). Absorption changesof b-type cytochrome induced by illumination were in just theopposite directions for oxygen- and nitrate-oxidized cells;the cytochrome was reduced in oxygen-oxidized cells and oxidizedin nitrate-oxidized cells. Antimycin enhanced the reductionand inhibited the oxidation, but had no inhibitory effect onthe oxidation of b-type cytochrome by nitrate. Dithionite-reducedminus ferricyanide-oxidized difference spectra of cells at 77?Kshowed two b-type cytochrome components with a bands at 556.5and 562 nm. The proportion of the b-562 component decreasedin cells grown under denitrifying conditions. It was concludedthat a b-type cytochrome is involved in the nitrate reduction.The b-type cytochrome was presumed to be an alternative to thecytochrome b in the photosynthetic cyclic electron transport. ¹ Present address: Japanese Red Cross Tokyo-to Komagome BloodCenter, Komagome 2-2-2, Toshima-ku, Tokyo 170, Japan. (Received August 13, 1981; Accepted December 5, 1981)     

Relation Between the Anatomy of the Testa, Water Permeability and the Presence of Phenolics in the Genus Pisum

The seed coats of Pisum elatius, P. fulvum and P. humile areimpermeable to water while those of P. sativum and P. humilex P. sativum are permeable. The anatomical structure of theseed coats and the location of phenolics and quinones in thecells is described. The barriers to permeation of water in theimpermeable seeds are the continuous, very hard, pectinaceouslayer of the caps of the palisade cells and the presence ofquinones in either the palisade or osteosclereid cells, in acontinuous layer of these cells. In water permeable seeds thecaps are looser and quinones discontinuous or absent in palisadeor osteosclereid cells. Pisum, testa, water permeability, quinones, phenolics, palisade cells, osteosclereids, pectinaceous caps     

Transient Gene Expression in Plant Cells Mediated by Agrobacterium tumefaciens: Application for the Analysis of Virulence Loci

A simple system is described for detection of the transfer ofT-DNA from Agrobacterium cells to suspension-cultured tobaccoBY-2 cells. A modified reporter gene for rß-glucuronidase(GUS) that contained an intron sequence was introduced intothe T-DNA region such that the GUS protein could be synthesizedin plant cells only after transfer of the T-DNA to plant nuclei.When BY-2 cells were co-cultured with Agrobacterium cells thatcontained the modified reporter gene, transient synthesis ofGUS protein was observed between 36 and 48 h after the onsetof co-culture. The level of GUS activity reached a plateau withinas little as 48 h. This temporal profile of GUS activation suggeststhat the transient activity might have been due to expressionof the GUS gene in the T-DNA that had been transferred to theplant nuclei but had not yet been integrated into the plantchromosomes. Levels of transient GUS activity were also examinedwith various vir mutants of Agrobacterium and in a mutant withan altered chromosomal acvB gene, the gene for a protein thathas been postulated to function outside bacterial cells. Duringco-culture with virB, virD2, virD4 and acvB mutants, GUS activityremained at background levels, and the GUS activity in the caseof the virE2 mutant was thirty-fold lower than with the wildtype. On the basis of these results, we discuss the roles ofthese genes during infection by Agrobacterium of plant cells. ⁴Present address: Biochemistry Laboratory, Kanebo Ltd., 5-3-28Kotobuki-cho, Odawara, Kanagawa, 250 Japan     

The Effects of Light Irradiation on the Orientation of Microtubules in Seedlings of Avena sativa L. and Pisum sativum L.

The effects of light irradiation on the arrangement of corticalmicrotubules (MTs) were examined in etiolated A vena mesocotylsand coleoptiles, and in Pisum epicotyls. Elongation of A venamesocotyls ceased as a result of irradiation with white lightwithin 1 h. The predominant orientation of MTs became more longitudinalwithin 1 h in epidermal cells and changed from transverse tooblique, after the elongation ceased, in parenchymal cells.Irradiation with red and with blue light also caused cessationof cell elongation and the same changes in the orientation ofMTs. Elongation of Avena coleoptiles ceased as a result of irradiationwith white light within 24 h. The predominant orientation ofMTs became more longitudinal in epidermal cells and changedfrom transverse to oblique in parenchymal cells. The changein orientation of MTs in epidermal cells preceded that in parenchymalcells. In Pisum epicotyls, elongation ceased as a result ofirradiation with white light within 1 h. Although the orientationsof MTs in epidermal cells did not show any remarkable change,those in parenchymal cells changed from transverse to obliqueafter cell elongation ceased. The change in orientation of MTs and the cessation of cell elongationof A vena mesocotyls induced by white-light irradiation wereboth significantly retarded by treatment with IAA. This resultsuggests that IAA is involved in maintaining the transverseorientation of MTs in Avena mesocotyls. (Received February 22, 1989; Accepted August 2, 1989)     

A Study of 'Phloeotracheids' in Haustoria of Santalaceous Root Parasites using Scanning Electron Microscopy

Tracheary elements within haustoria of six santalaceous rootparasites have been examined by scanning electron microscopywith particular reference to those cells which contain granules.These cells have been called ‘phloeotracheids’ byearly workers. The granules lie free within the lumen of thecell from which a protoplast is absent at maturity. The granulesoccur in short vessel elements and in imperforate cells whichare thought to represent a specialized type of tracheid. Phloeotracheidsare confirmed in Exocarpus bidwilliiand Mida salicifolia andreported for the first time in Buckleya distichophylla, Comandraumbellata, Nestronia umbellataand Pyrularia pubera. The structureof phloeotracheids shown by scanning electron microscopy isdiscussed in relation to previous studies.     

中华稻蝗消化道内分泌细胞的鉴别与定位

采用整块组织Grimelius银染法和过氧化物酶标记的链霉亲和素免疫组织化学技术,结合生物统计学分析,对中华稻蝗Oxya chinensis消化道内分泌细胞进行鉴别与定位。结果表明：嗜银细胞分布于中华稻蝗的胃盲囊、中肠和后肠各段,以中肠和直肠中最多（P<0.05）, 前肠中未见分布。免疫组织化学法检测出了五羟色胺（5-hydroxytryptamine, 5-HT）、 胃泌素（gastrin, Gas）、 胰高血糖素（glucagon, Glu）和胰多肽（pancreatic polypeptide, PP）细胞, 未检出生长抑素（somatostatin, SS）细胞。免疫阳性细胞分布于中肠和后肠中, 前肠中未见分布。5-HT细胞和Gas细胞均主要分布于胃盲囊、中肠及直肠中,且均以直肠中最多（P<0.05）。Glu细胞在胃盲囊及整个中、后肠均有分布, 在中肠和直肠中最多（P<0.05）。PP细胞主要分布于中肠、回肠和直肠中,中肠中分布密度最大（P<0.05）。本研究显示中华稻蝗消化道中存在多种内分泌细胞,它们的分布情况与其他节肢动物相比存在一定的共性,也有其一定的特异性,可能与中华稻蝗特定的消化道结构和消化生理功能有关。     

Studies on the Development of the Air Pores and Air Chambers of Marchantia paleacea: 1. Light Microscopy

The ontogeny of the air pores and air chambers of Marchantiapaleacea begins with the schizogenous development of protodermalintercellular spaces of the initial apertures, and is completedwith the formation of the air pores and giant sub-epidermalair chambers bearing numerous photosynthetic filaments. Intercellularspace formation commences from the thallus surface and proceedsinwards to the first internal layer of cells. The cells amongwhich spaces develop do not originate from one mother cell.Spaces are formed only in the regions of the intersection ofthe anticlinal walls of three, four, or sometimes more successivederivatives of S₁, S₃ and S₄ segments of the apical cell, oneor two of which have been divided periclinally and the restanticlinally. Protodermal intercellular spaces appear in mostor all the corners of these cells, the anticlinal walls of whichexhibit an opposite disposition. The S₁, S₂, S₃ and S₄ segmentsare produced by definite divisions of a five-sided apical celland by a series of divisions give rise to initial cells of theinternal layers of the thallus and initial cells of the protodermaland sub-protodermal layers. The concept of a quiescent apicalcell cannot be accepted, since dividing apical cells have beenobserved, and the pattern of wall disposition of the thallusapex cannot be explained without the active participation ofthe apical cell. The air chambers are apparently of exogenous origin. They resultfrom the broadening of the bottom of the initial apertures bythe coordination of the rate of anticlinal divisions and growthof the protodermal and sub-protodermal cells surrounding theintercellular spaces of the initial apertures. The ontogenyof the pore rings starts at an advanced stage of air chamberformation not from a mother cell but from the cells which surroundthe closed entrance of the air chamber, by a shift of the planeof division from anticlinal to periclinal. Before the periclinaldivisions a new axis of growth perpendicular to the thallussurface is established in the mother cells of the pore. By a polarized growth into the air chamber followed by periclinaldivisions, the cells of the floor form initial cells of thephotosynthetic filaments. The latter divide again to form singleor branched photosynthetic filaments. Marchantia paleacea, air pore, air chamber     

Endogenous Levels of Gibberellins, IAA and Cytokinins in Catharanthus Crown Gall Tissues of Different Tumor Types

Endogenous levels of gibberellins, auxins and cytokinins incrown gall tissues of Catharanthus roseus G. Don. (Vinca roseaL.) of two different tumor types, induced by a nopaline-typeand an octopine-type Ti-plasmid, were examined. The levels of gibberellins were higher in nopaline-type V208cells than in octopine-type V277 cells. GA₁₂ and GA₂₄ were identifiedfrom V208 cells. The level of IAA was 20-fold higher in V208cells than in V277 cells. The level of trans-ribosylzeatin washigher in V277 cells than in V208 cells, whereas trans-zeatinwas present at higher levels in V208 cells than in V277 cells. Our results revealed that the production of gibberellins, aswell as of IAA and cytokinins, in the crown gall tissues ofCatharanthus roseus differed between the octopine- and nopaline-typetumors, even though the crown galls apparently grow as unorganizedaggregates of cells in both cases. ⁴Present address: Facultad de Ciencias, Pontificia UniversidadJaveriana, Bogota, Colombia. (Received September 29, 1989; Accepted February 2, 1990)     

Arrangement of Cortical Microtubules in Avena Coleoptiles and Mesocotyls and Pisum Epicotyls

Cortical microtubules (MTs) in coleoptiles and mesocotyls ofAvena sativa and epicotyls of Pisum sativum were examined byimmunofluorescence. In elongating Avena coleoptiles whose elongationis less localized, the orientations of cortical MTs of parenchymaand adaxial epidermal cells, and abaxial epidermal cells aretransverse, and oblique or longitudinal, respectively, and doesnot differ between the upper, middle and lower parts. The transverseMTs in parenchyma and adaxial epidermal cells turns to obliqueor longitudinal ones after elongation stops. The obliquity ofMTs in abaxial epidermal cells also tends to become steeperas elongation comes to a stop. In Avena mesocotyls and Pisumepicotyls whose elongation is localized, the orientation ofcortical MTs of cortical cells in the elongating region is relativelytransverse. The epidermis has intermingling cells of transverseor oblique MTs. In the non-elongating region, MT orientationbecomes steeper both in the cortex and epidermis. The present results indicate that whatever the degree of localizationof the elongation, the obliquity of MTs in these organs is steeperin epidermal than in inner tissue cells and becomes steeperas elongation stops in both tissues. (Received October 26, 1987; Accepted April 19, 1988)     

Root Gravitropism in a Cultivar of Zea mays Whose Columella Cells Contain Starch-deficient Amyloplasts

Starch occupies 4.2 per cent of the volume of plastids in calyptrogencells in primary roots of Zea mays L. cv. vp-7 wild type. Plastidsin calyptrogen cells are distributed randomly around large,centrally located nuclei. The differentiation of calyptrogencells into columella cells is characterized by cellular enlargementand the sedimentation of plastids to the bottom of the cells.Although sedimented plastids in columella cells do not containsignificantly more starch than those in calyptrogen cells, primaryroots are graviresponsive. The onset of root gravicurvatureis not associated with a significant change in the distributionof plastids in columella cells. These results indicate thatin this cultivar of Z. mays (1) the sedimentation of plastidsin columella cells is not based upon their increased densityresulting from increased starch content alone, (2) starch-ladenamyloplasts need not be present in columella cells for rootsto be graviresponsive, and (3) the onset of root gravicurvaturedoes not require a major redistribution of plastids in columellacells. Columella cell, gravitropism (root), plastids, root cap, Zea mays     

Developmental Ultrastructure of Cells and Plastids in the Petals of Wallflower (Erysimum cheiri)

An ultrastructural study of petal cells of wallflower (Erysimumcheirii) of the family Brassicaceae shows that the adaxial epidermalcells are of the conical papillate type whereas the cells ofthe abaxial epidermis are lenticular in shape. The abaxial epidermiscontains stomata, which are solitary and lack any obvious subsidiarycells. Pigmentation is apparent in both epidermal and internalmesophyll cells and results from the presence of both chromoplastsand large cytoplasmic vesicles containing pigment. These pigmentedvesicles are very obvious in preparations of fixed isolatedpetal cells. Chromoplasts are of the globular type and are presentin significant numbers in both epidermal and mesophyll cells.Division of chloroplasts in young petals prior to bud breakappears to give rise to the populations of chromoplasts observedin mature petals since there was no evidence of chromoplastdivision itself. The development of wallflower petals and theirchromoplasts is discussed in relation to development of petalsin the related species Arabidopsis thaliana. Copyright 1999Annals of Botany Company Wallflower, Erysimum cheiri, Chieranthus, petal development, chromoplasts, chloroplast differentiation.