Histochemistry and Senescence of Colleters of Allamanda cathartica (Apocynaceae)

Allamanda have colleters on the adaxial bases of petiole, bractsand bracteoles. Each colleter is finger shaped, differentiatedinto a long head on a short stalk. Vasculature to the colleteris absent but branched laticifers are present among the centralcells. Secretion of the colleter contains glucose and rhamnose.The major elements present in the exudate are Na, Fe and Zn.Starch, protein and lipids were identified, with lipids themajor component. Petiolar colleters are persistent and becomestiff due to the lignification occurring in the walls of epithelialand central cells. Because of over-lignification in the centralcells, the cell lumen is highly reduced. Allamanda, colleter, laticifer, secretion, senescence     

A Cytochemical Study of Differentiation and Breakdown of Vessel End Walls

A cytochemical approach was used to study the architecture ofthe end wall and its modifications during vessel differentiationin Populus italica and Dianthus caryophyllus. The combinationof cytochemical techniques with selective extractions to removeend wall subunits provides information on the different componentsof the end wall and on the method of perforation. The end wallappears to be formed mainly from pectins and hemicelluloses.No cellulosic components nor lignin were found though the 3,3'-diaminobenzidine assay revealed the presence of peroxidases.That the method of perforation may depend on the species isdiscussed. Dianthus caryophyllus L., carnation, Populus italica, poplar, xylem vessel, cell wall, cytochemistry, vessel differentiation     

From the sap's perspective: The nature of vessel surfaces in angiosperm xylem

Premise of the Study

Xylem sap in angiosperms moves under negative pressure in conduits and cell wall pores that are nanometers to micrometers in diameter, so sap is always very close to surfaces. Surfaces matter for water transport because hydrophobic ones favor nucleation of bubbles, and surface chemistry can have strong effects on flow. Vessel walls contain cellulose, hemicellulose, lignin, pectins, proteins, and possibly lipids, but what is the nature of the inner, lumen‐facing surface that is in contact with sap?

Methods

Vessel lumen surfaces of five angiosperms from different lineages were examined via transmission electron microscopy and confocal and fluorescence microscopy, using fluorophores and autofluorescence to detect cell wall components. Elemental composition was studied by energy‐dispersive X‐ray spectroscopy, and treatments with phospholipase C (PLC) were used to test for phospholipids.

Key Results

Vessel surfaces consisted mainly of lignin, with strong cellulose signals confined to pit membranes. Proteins were found mainly in inter‐vessel pits and pectins only on outer rims of pit membranes and in vessel‐parenchyma pits. Continuous layers of lipids were detected on most vessel surfaces and on most pit membranes and were shown by PLC treatment to consist at least partly of phospholipids.

Conclusions

Vessel surfaces appear to be wettable because lignin is not strongly hydrophobic and a coating with amphiphilic lipids would render any surface hydrophilic. New questions arise about these lipids and their possible origins from living xylem cells, especially about their effects on surface tension, surface bubble nucleation, and pit membrane function.     

Isolation and structure of 13,18-dehydroexcelsin,a quassinoid,and glaucarubol from Ailanthus excelsa

A new quassinoid, 13,18-dehydroexcelsin and glaucarubol have been isolated from the bark of Ailanthus excelsa.     

Cellulolytic Activities of Botryodiplodia theobromae Pat

Botryodiplodia theobromae Pat., a ‘stain’ fungusisolated from stained and decaying wood of Bombax buonopozenseP. Beauv., attacked isolated blocks of Bombax wood. In theseit perforated the cell walls and formed elongated cavities withpointed ends, usually in chains, in the secondary wall. Thefungus attacked cellulose in a similar manner to soft rot fungi.It used starch and other saccharides present in the wood ofBombax buonopozense as initial substrates before degrading thecellulose and hemicellulose components of the wood. It did notdegrade the lignin component of the wood. Cellulase, ß-glucosidase, and amylase activities weredetected in culture filtrates of Botryodiplodia theobromae.Cellulase synthesis was delayed when the wood powder in themedium contained starch and saccharides. Amylase, too, behavedlike an inducible enzyme and disappeared from the cultures onexhaustion of starch. Electron microscopy showed that cellulose extracted from thewood of Bombax buonopozense was fragmented into small particleson treatment with the culture filtrates of Botryodiplodia theobromae.Cellobiose, glucose, and another saccharide, probably cellotriose,were identified as the products of enzymatic hydrolysis of cellulose. These results are discussed in relation to the attack and degradationof the wood of Bombax buonopozense by Botryodiplodia theobromaein nature.     

The Composition of Phloem Exudate and Xylem Sap from Tree Tobacco (Nicotiana glauca Grah.)

The composition of xylem sap and exudate from stem incisionsof Nicotiana glauca Grah. was compared in detail. Exudationfrom stem incisions occurred over a 5 min period in certainplants, enabling collection of 5–30 µl of sap. Therate of exudation showed an exponential decline. Exudate hada high dry matter content (170–196 mg ml^–1) andhigh sugar (sucrose) levels. Xylem sap had a low pH (5.8) andexudate a pH of 7.9. Glutamine dominated the amino compoundsin xylem sap and exudate, and K⁺ was the major cation. Totalamino compounds in stem exudate reached 10.8 mg ml^–1 whereasxylem sap contained much lower levels (0.28 mg ml^–1).All mineral elements and amino compounds with the exceptionof calcium were more concentrated in stem exudate than in xylemsap. Sucrose was labelled heavily in stem exudate following pulsingof an adjacent leaf with ¹⁴CO₂. A concentration gradient ofsugar (2.1 bar m^–1) was recorded for stems. Levels ofsucrose, amino compounds and K⁺ ions in stem exudate showeda diurnal periodicity. Each commodity reached maximum concentrationat or near noon and minimum concentration about dawn. The evidencesuggests that exudate from stem incisions of N. glauca is arepresentative sample of solutes translocated in the phloem. Nicotiana glauca Grah., phloem sap, xylem sap, sucrose, amino compounds, mineral ions     

The Distribution of Silicon Deposits in the Roots of Molinia caerulea (L.) Moench. and Sorghum bicolor (L.) Moench.

The occurrence of silica in relation to meristematic zones andthe thickening of the endodermis in the roots of Molinia caerulea(L.) Moench. and Sorghum bicolor (L.) Moench. has been investigatedby means of the electron-probe microanalyser and the scanningelectron microscope. In proximal regions of mature roots ofM. caerulea, the central strengthening tissue of the stele,the vessel walls, the endodermis and the sub-epidermal sclerenchymaare areas of heavy accumulation. The distal regions of suchroots are relatively free of silicon and show little thickeningof the inner tangential walls of the endodermis or of the cellsof the strengthening tissues. The thickening of these elementsis shown to be associated with their location and the age ofthe root. In the proximal regions of S. bicolor, silicon is detected andlargely confined to the inner tangential walls of the endodermiswhich display some thickening. In addition, discrete and evenly-distributeddeposits varying in size partly fill the lumen of this layer.Some cells exhibit a number of smaller protrusions. High magnificationsof these lumen deposits show a distinct granular structure incontrast to the very uniform pattern of the wall deposits. The results are compared with deposits in grass leaves and inflorescencebracts and in woody perennials. The presence of silicon in additionto suberin, lignin and polyphenols in the thickened endodermalwall is also discussed in relation to the recognized functionof the endodermis.     

3S,24S,25-Trihydroxytirucall-7-ene from Ailanthus excelsa

A new triterpene has been isolated from the root bark of Ailanthus excelsa (Simaroubaceae) and identified as 3S,24S,25-trihydroxytirucall-7-ene.     

Functional Anatomy of the Ovule in Broad Bean (Vicia faba L.): Ultrastructural Seed Development and Nutrient Pathways

Ovules of broad bean (Vicia faba L.) were studied to discloseultrastructural features, which can facilitate nutrient transportto the embryo sac from 10 d after pollination (DAP) to the matureseed. Fertilization occurs during the first 24 h after pollination.The endosperm is a coenocyte, which is eventually consumed bythe embryo. By 10 DAP the inner integument is degraded and theouter integument adjoins the embryo sac boundary. The heart-shapedembryo approaches the embryo sac boundary at two sites, whichhere are named contact zones. Small integument cells in theneighbourhood of the first formed contact zones become separatedby prominent intercellular spaces. A heterogenous scatteringmaterial, probably representing secretion products accumulatesin these spaces. By 14-16 DAP the integument exudate disappears,and the suspensor degenerates. As the contact zones increasein size, wall ingrowths form a bridging network in the narrowspace between the embryo sac boundary and the extra-embryonicpart of the endosperm wall. The epidermal cells of the embryoseparate adjacent to these zones, and develop conspicuous wallingrowths. At 20 DAP vacuoles showing various stages in formationof protein bodies appear in the cells of the embryo.Copyright1994, 1999 Academic Press Vicia faba, broad beans, ovule, seed, nutrient transport     

The Structure and Cytochemistry of the Pistil of Sternbergia lutea (Amaryllidaceae)

Studies were carried out on structural and cytochemical aspectsof the pistil of Sternbergia lutea (L.) KerGawl. The stigmais of the wet papillate type; the papillae are unicellular andare arranged densely around the rim of a funnel-shaped stigma.The stigma exudate is limited and is confined to the bases ofthe papillae and the inner lining of the stigma. The papillaeare smooth in the distal part and are covered with intact cuticle-pelliclelining. The cuticle is disrupted at places towards the baseof the papillae releasing the exudate. The exudate is rich inpectins and other polysaccharides but poor in proteins and lipids.The papillae show dense cytoplasmic profiles with extensiveendoplasmic reticulum (ER), abundant mitochondria, polyribosomesand active dictyosomes. The style is hollow. The stylar cavityis surrounded by two to four layers of glandular cells. In theyoung pistil the canal is lined with a continuous cuticle, butin the mature pistil the cuticle becomes disrupted and the canalis filled with the secretion produced by the cells of the surroundingglandular tissue. Ultrastructurally, the cells of the glandulartissue are very similar to the stigmatic papillae. The innertangential wall of the cells bordering the canal is uniformlythicker than other walls. The secretion in the stylar canal,as well as the intercellular spaces of the glandular tissue,stain intensely for pectins and polysaccharides but poorly forproteins and lipids. Pollen tubes grow through the stylar canal.Structural and cytochemical details of the pistil of Sternbergiaare compared with other hollow-styled systems. Pistil, Sternbergia lutea (L.) Ker-Gawl., stigma and style, structure and cytochemistry     

Studies in the Physiology of Parasitism: XXI. The Production and Properties of Pectic Enzymes secreted by Fusarium moniliforme Sheldon

Fusarium moniliforme secreted macerating enzymes in liquid mediaonly when these contained certain natural extracts, pectic substances,or galacturonic acid. Apple extract was unsuitable for enzymesecretion and also inhibited enzyme secretion in synthetic mediaotherwise suitable. Protopectinase activity of solutions was highest in the pH range8·0–9·0, was rapidly lost at temperaturesabove 50–60° C., and was reduced by concentrationsof phosphate higher than 0·02 M. The enzyme was partiallypurified by precipitation in 60 per cent. acetone at pH 6·0. Protopectinase solutions also contained an enzyme which reducedthe viscosity of solutions of various pectic substances. Theproperties of this enzyme were, in general, similar to thoseof protopectinase. When activity of enzyme solutions was measured by the liberationof reducing groups, pectate solutions were more rapidly degradedthan were solutions of a high methoxyl pectin, particularlyin the early stages of the reaction. Paper chromatography ofthe products formed showed that pectate and pectin were degradedin different ways. Although the pathogen readily secreted protopectinase in potatoextract, potato tubers were not readily parasitized. In contrast,Fusarium avenaceum which readily attacked tubers, secreted littleprotopectinase in potato extract.     

Developmental Aspects of Ultrastructure, Histochemistry and Receptivity of the Stigma of Nicotiana sylvestris

The bilobed papillate stigma of Nicotiana sylvestris Speg. andComes, is covered at maturity with a copious exudate containinglipid, protein and carbohydrate. The stigma is receptive fromthe very early stage of development and it also stains positivelyfor esterase activity. The stigma has three distinct zones:an epidermis with papillae; a subepidermal secretory zone; anda parenchymatous ground tissue. The behaviour of the cells ofthese three zones has been followed from 6 d before anthesisto one day after anthesis and pollination. The cells of theepidermis and the secretory zone stain intensively for lipids,proteins and carbohydrates in the initial stages. The secretoryzone develops large intercellular spaces containing heterogenoussecretory products which also stain positively for the aforesaidthree compounds. At maturity the secretory products are releasedto the surface through gaps formed in the epidermis by cellseparation. The main secretion of the stigma is produced bythe cells of the secretory zone. Less secretion is derived fromthe stigmatic papillae. Some amount of secretion is also releasedfrom the stylar transmitting tissue adjoining the stigma. Theglandular cells of the stigma contain numerous plastids, mitochondria,ribosomes, ER, cytoplasmic lipid droplets and some dictyosomes.The plastids and the vacuoles in the secretory cells of thestigma have a lot of electron dense (osmiophilic) inclusionsrespectively in the initial and later stages of development.The former are probably involved in the production of thesematerials. It is suggested that the proteins are directly secretedby rough ER compartments whereas smooth ER is involved in thesynthesis of lipidic materials. The carbohydrate moiety of theexudate is released by the eccrine mode (sugar mono- and dimers)with some addition of polymers by disintegration of the middlelamellae. The means by which the lipidic and osmiophilic materialis extruded remains unclear. Nicotiana sylvestris, stigma receptivity, organization, stigmatic secretory system, stigmatic exudate     

Flower-Inducing Activity of Phloem Exudates from Pharbitis cotyledons Exposed to Various Photoperiods

The phloem exudate prepared from the cotyledons of Pharbitisseedlings that had been exposed to a single dark period (oflonger than 10 h) induced flowering in cultured apices excisedfrom non-induced seedlings. The flower-inducing activity ofthe exudate increased as the seedlings were exposed to longerperiods of darkness. The highest activity was associated withthe exudate taken from cotyledons exposed to a single 16-h darkperiod. The activity of the exudate taken from cotyledons exposedto an inductive dark period was clearly reduced by interruptionof the dark period. The addition of exudate taken from threecotyledons to 10 ml of medium resulted in the highest flower-inducingactivity. About 50% of cultured apex explants formed floralbuds, even when the concentration of the exudate was reducedto 0.1 cotyledon equivalents per 10 ml of medium. The flower-inducingactivity of the exudate appeared to be heat-stable. (Received December 13, 1991; )     

Pectolytic Activity by the Haustorium of the Parasitic PlantOrobancheL. (Orobanchaceae) in Host Roots

The possible involvement of enzymes in the penetration of intrusivecells of the parasitic angiospermOrobancheinto host root tissueswas studied using cytochemical and immunocytochemical methods.Pectin methyl esterase (PME) was detected, with specific antibodies,in the cytoplasm and cell walls ofOrobancheintrusive cells andin adjacent host apoplast. Depletion and chemical changes ofpectins in host cell walls were shown by histochemical stainingwith PATAg, which detects carbohydrates that are sensitive toperiodic acid, especially pectins, and with the monoclonal antibodiesJIM 5 and JIM 7 that label pectins with low and high rates ofesterification, respectively. Galacturonic sequences with lowrates of esterification were more abundant in host cell wallsadjacent to the parasite, which is consistent with pectin de-methylationby PME release from the parasite. Pectins were absent in middlelamellae and in host cell walls neighbouring mature intrusivecells of the parasite, consistent with further degradation ofpectins by other enzymes. These results provide the first directevidence for the presence and activity of a pectolytic enzymein the infection zone of the haustorium of a parasitic angiosperminsitu.Copyright 1998 Annals of Botany Company Broomrape;Orobanche; parasitic weed; haustorium; pectin methyl esterase; pectin; cell wall.     

Regeneration of the peripheral nerves in the lumen of implanted vessels

Regeneration of the rat peripheral nerve in the lumen of implanted aorta was examined over 4 months. The nerve invaded the vessel with its entire trunk and went out of the distal vessel to the adjacent muscles. Inflammatory infiltrations were observed only outside the implant. Only insignificant number of inflammatory elements were inside the lumen. These conditions favored good regeneration of the nerve without forming a neuroma at the end of the cut nerve. The regenerating myelinized fibers formed spiral-like or bush-like terminals on muscle fibers. The model of directed regeneration of the peripheral nerves in the vascular lumen holds promise for rapid and accurate reinnervation of tissues and organs.     

益智胚珠的珠心冠原与承珠盘细胞壁的组织化学研究

用组织化学方法研究了益智胚珠中珠心冠原与承珠盘细胞壁的组成。珠心冠原细胞壁含有纤维素、胼胝质、果胶质,但不含栓质。承珠盘细胞壁含有纤维素、木质素、果胶质,也不含栓质。讨论了珠心冠原与承珠盘细胞壁的组成及承珠盘的可能功能。     

An Ultrastructural Investigation of Protein and Lipid Reserves in the Endosperm of Pinus pinea L.

Proteins and lipids are the two main storage substances in theendosperm of Pinus pinea L. In imbibed seeds, lipid and proteinbodies almost completely fill the cell volume and only a fewpoorly structured organelles can be detected. During germinationthe storage substances are reabsorbed whilst plastids, roughendoplasmic reticulum, ribosomes and polysomes, microbodies(glyoxysomes, most likely) become evident. Starch is also formed.When the cotyledons emerge completely from the endosperm envelopethe latter shows highly vacuolate cells but still rich in cytoplasmicorganelles, lipids, protein bodies and starch grains. Different patterns of protein body degradation are discussed. Pinus pinea, endosperm, ultrastructure, protein, lipid, stored reserves     

The Nature of the Membrane Around the Egg of Peteridium aquilinum (L.) Kuhn

The mature egg of Pteridium aquilinum is surrounded by an acetolysis-resistantmembrane. Cytochemical tests indicate that this consists oflipid, possibly in a polymerized form. There is no evidencefor the presence of polysaccharide, protein, lignin, suberin,or sporopollenin. The membrane is believed to be formed frommaterials secreted by the egg itself, and to be analogous tothe inner portion of the megaspore membrane of a gymnospermwhich has a comparable origin and staining properties.     

Decay rate and substrate quality of fine roots and foliage of two tropical tree species in the Luquillo Experimental Forest,Puerto Rico

Decomposition rates, initial chemical composition, and the relationship between initial chemistry and mass loss of fine roots and foliage were determined for two woody tropical species, Prestoea montana and Dacryodes excelsa, over a gradient of sites in two watersheds in the Luquillo Experimental Forest, Puerto Rico. At all locations, fine roots decayed significantly more slowly than foliage during the initial 6 months.Substrate quality of the initial tissue showed marked differences between roots and foliage when using cell wall chemistry, secondary chemistry and total elemental analysis as indices. Quantity of acid detergent fiber (ADF) (non-digestible cell wall fiber) and lignin content were higher for roots than leaves: D. excelsa roots had 55.3% ADF and 28.7% lignin while leaves had 36.2% ADF and 11.8% lignin; P. montana roots had 68.0% ADF and 26.8% lignin while leaves had 48.5% ADF and 16.1% lignin. Aluminum concentrations were higher in fine roots (843 mg kg^–1 in D. excelsa, 1500 mg kg^–1 in P. montana) than leaves (244 mg kg^–1 in D. excelsa, 422 mg kg^–1 in P. montana), while calcium concentrations were higher in foliage (5.5 mg g^–1 in D. excelsa, 7.8 mg g^–1 in P. montana) than roots (3.4 mg g^–1 in D. excelsa, 3.1 mg g^–1 in P. montana). Nitrogen did not show any trend with tissue or species type. A linear model between mass remaining after 6 months and initial tissue chemistry could be developed only for calcium (r²=0.64).     

Structural Aspects of Cluster Root Development and their Possible Significance for Nutrient Acquisition in Grevillea robusta (Proteaceae)

Light microscopy (LM), scanning (SEM) and transmission electronmicroscopy (TEM) were used to study structure and function ofcluster roots inGrevillea robusta . These roots were developedduring growth ofG. robustaseedlings in modified Hoagland's solutionlacking phosphate. Cluster rootlets formed root hairs, basipetally,only after completing their determinate development. The rootlethairs branched in two ways and some had apical swelling. Rootletswith hairs produced two different forms of exudate, one fibrousand the other globular in nature. The fibrous material appearedto be synthesised in the cortical cells. It is released by exocytosisfrom the epidermis. Rootlet hairs produced only fibrous exudate.They attached firmly to pieces of vermiculite. The significanceof cluster roots is discussed within the context of patchy soilresources. Grevillea robusta ; Silky oak; Proteaceae; cluster roots; morphology; mucilage; nutrient acquisition; root exudates