Sequence Analysis of the Bacillus subtilis 168 Chromosome Region between the sspC and odhA Loci (184{degrees}-180{degrees})

The nucleotide sequence of 45,389 bp in the 184°-;180°region of the Bacillus subtilis chromosome, containing the cgecluster, which is controlled by the sporulation regulatory proteinGerE, was determined. Fifty-four putative ORFs with putativeribosome-binding sites were recognized. Seven of them correspondto previously characterized genes: cgeB, cgeA, cgeC, cgeD, cgeE,ctpA, and odhA. The deduced products of 25 ORFs were found todisplay significant similarities to proteins in the data banks.We have identified genes involved in detoxification, cell walls,and in the metabolism of biotins, purines, fatty acids, carbohydratesand amino acids. The remaining 22 ORFs showed no similarityto known proteins. Both an attachment site of the SPßprophage and 2 new putative DNA replication terminators wereidentified in this region.     

Cell Wall Autolysis During Kiwifruit Development

Cell walls prepared from developing kiwifruits showed autolyticactivity. The proteins extracted from active walls were alsoable to release carbohydrates from inactive cell walls. Analysisof the sugars released, using both procedures, showed that uronicacids were the major component, especially during the firsthours of incubation, although neutral sugars such as glucoseand galactose were also present. Most of the carbohydrates autolyticallyreleased from the cell wall eluted in the void volume on a BioGel P2 column. However, carbohydrates released from inactivecell walls by the protein extract mostly eluted in the monosaccharideuronic acid and glucose peaks. The autolytic activity of isolatedcell walls, as well as the glycosylhydrolase activity of theproteins extracted from the cell walls, showed important changesduring fruit development. The differences between autolyticactivity and the glycosylhydrolase activity against the cellwall suggest that the glycosylhydrolases ‘in muro’are subjected to some regulatory mechanism which disappearswith their extraction. Finally, the role of glycosylhydrolases,such as polygalacturonases and galactosidases, in relation tocell wall changes in fruits, is discussed.Copyright 1998 Annalsof Botany Company Actinidia deliciosa; autolysis; cell wall enzymes; fruit growth; glycosylhydrolases; kiwifruit.     

Changes in the composition of exocellular proteins of suspension-cultured Lupinus albus cells in response to fungal elicitors or CuCl2

Among many aspects of plant defence responses to pathogenicinfection are changes in the composition of the exocellularmatrix. To study potential defences in white lupin (Lupinusalbus L.), suspension-cultured cells were treated for 24 h withone of three different elicitors: CuCl₂, and two fungal elicitorpreparations from purified cell walls of yeast and ColletotrichumIindemuthianum. Two subsets of exocellular proteins: ionicallyboundwall proteins and proteins secreted into culture medium, wereisolated, and their patterns compared following electrophoreticseparation. Only a few proteins were observed in culture filtrateswith dominating bands at 27, 33, and 42 kDa. About 30 proteinswere observed in cell wall extracts. Changes in protein intensitiesevoked by elicitor treatments depended on the type of elicitorused, the age and composition of lupin cell culture, and concentrationof applied fungal elicitor. Based on these observations, tenproteins were chosen for N-terminal sequencing, and sequences5–30 amino acids long for nine proteins were obtained.Three of the major proteins sequenced were identified as acidicexocellular chitinase, polygalacturonaseinhibiting protein,and germin/oxalate oxidase. Key words: Lupinus aibus, defence response, exocellular proteins, elicitation, N-terminal amino acid sequencing, suspension culture     

Glycanases Associated with Cell Walls of Cicer arietinum L: Arabinogalactan Degradation

The proteins extracted from Cicer arietinum cell walls showedendo-glycanase activity against the water-soluble hemicellulosicpolysaccharides extracted with 4% KOH. Such endo-glycanase activitywas able to shift the molecular mass distribution of the arabinogalactancausing a decrease in its average molecular mass, as well asto release small oligosaccharides constituted exclusively ofarabinose. Exo-glycanase activities such as glucosidase andgalactosidase were also present. Key words: Arabinogalactan, cell wall, Cicer arietinum, glycanase, hemicellulose     

Expression of Cell Wall Proteins in Seeds and During Early Seedling Growth of Araucaria araucana is a Response to Wound Stress and is Developmentally Regulated

Araucaria araucana seeds and seedlings respond to wounding after48 h with a 3- to 4-fold increase of hydroxyproline-rich glycoproteins(HRGP) in the cell walls of the embryo and with a 15-fold increasein the cell walls of the megagametophyte. The megagametophytewalls accumulate six times more hydroxyproline per µgof cell wall protein than the embryo in this wound response.Tissue immunoprints of different parts of seeds and seedlingsobtained with polyclonal antibodies raised against HRGP fromcarrot roots or soybean seed coats indicate that the responseis due to an increase in a protein similar to the ones seenin carrot roots or soybean seed coats. Western blots of embryoand megagametophyte cell wall proteins subjected to SDS-PAGEshow three bands that cross-react with these antibodies. Ina native cationic gel system followed by Western blot analysis,only two bands react with these antibodies. Expression of suchproteins in Araucaria araucana seeds seems to be developmentallyregulated and tissue specific, since they are present mainlyin the megagametophyte and the root cap of the embryo. Key words: Araucaria araucana, seeds, seedlings, cell walls, hydroxyproline-rich glycoproteins     

Fine Structure of Hydrolysed Primary Walls in Tracheary Elements of Petiolar Xylem in Eucalyptus delegatensis

The hydrolysed lateral primary walls of tracheary elements ofthe petiolar xylem of Eucalyptus delegatensis were examinedby electron microscopy. Vessel-vessel and vessel—tracheidhydrolysed walls were strikingly different in appearance fromtracheid—tracheid walls. The difference seemed to be inthe degree to which the primary walls were hydrolysed. The observationssuggest the wall hydrolysis to be an ordered and controlledprocess. Eucalyptus delegatensis, hydrolysed wall, petiolar xylem, tracheary elements     

Studies on Mature Seeds of Cuscuta pedicellata and C. campestris by Electron Microscopy

The seeds of Cuscuta pedicellata have been investigated by transmissionand scanning electron microscopy. Additional observations havebeen made on seeds of C. campestris by SEM only. The seed coatconsists of an outer single epidermis, two different palisadelayers, and an inner multiparenchyma layer. The outer epidermalwall in C. pedicellata has a thick cuticle and zones rich inpectic substances. The thicker ‘U-shaped’ cell wallsin the outer palisade layer are strengthened by a wall layerof hemicellulose. The inner palisade layer has thick walledcells with a ‘light line’. The inner cell wall ofthe compressed multiparenchyma layer has a thin cuticle. A fairlythick cuticle is positioned directly on the endosperm surface.The aleurone cell walls are different from the remaining endospermwalls. The latter are thick and believed to be of galactomannans.There is a ‘clear’ zone between the plasmalemmaand the cell wall in the aleurone cells. The embryo cells arepacked with lipids and proteins. In Cuscuta campestris mostendosperm has been absorbed during the seed development. Theembryo apex has two minute leaf primordia. The features of theCuscuta seeds are discussed in relation to functional and environmentalconditions. Cuscuta pedicellata, Cuscuta campestris, seed, seed coat, cuticle, cell walls, endosperm, aleurone cells, galactomannan, embryo, TEM, SEM     

Stability of the Cell Walls of Neurospora crassa during Autolysis

The influence of autolysis upon the cell walls of Neurosporacrassa has been studied. This fungus was grown at 24 °Cin agitated and aerated cultures in a synthetic medium during60 days. At convenient intervals samples of culture were taken,mycelium separated, and dried to constant weight. From aliquotsof these mycelia cell walls were prepared, dried, weighed, andanalysed for total nitrogen, phosphorus, amino acids, lipids,and protein. No changes in the chemical composition of the wallscould be detected. The percentage of walls continuously increasedduring autolysis. These results strongly suggest that cell wallsof N. crassa are unaffected by autolysis. Examination of thefine structure of the whole mycelium at different ages duringautolysis seemed to confirm these findings.     

Studies on the Cell Wall of Chlorella V. Comparison of the Cell Wall Chemical Compositions in Strains of Chlorella ellipsoidea

Cell walls of four strains of Chlorella ellipsoidea (IAM C-27,C-87, C-102 and C-183) were compared as to their chemical compositions.Many differences were found: (1) The sugar composition of alkali-soluble cell walls differedin quantity as well as quality with glucuronic acid being foundonly in C-27 and C-87. (2) In alkali-insoluble cell walls glucosamine was found onlyin C-27. The other three strains contained mainly glucose. (3) The amino acid compositions of the alkali-insoluble cellwalls markedly differed among the four strains. The cell wallof C-102 contained more amino acids than carbohydrates, butC-27 and C-87 contained extremely little amino acid. In addition to the variation in cell wall composition, the opticalanisotropy findings also differed for these cell walls of Chlorellastrains which had been grouped as the same species. (Received August 16, 1983; Accepted December 27, 1983)     

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.     

Silicon Deposition in the Roots of Hordeum sativum Jess, Avena sativa L. and Triticum aestivum L.

Electron-probe microanalysis was used to investigate the locationof silicon at the proximal end of the seminal and adventitiousroots, of almost mature field-grown specimens of Hordeum sativumJess., Avena sativa L. and Triticum aestivum L. In the seminal roots silicon was confined to the endodermis,where it was present in the thickened inner tangential and radialwalls. The outer tangential walls also contained silicon inall of the cells in wheat and in occasional cells in barleyand oats. The adventitious roots of the three cereals displayed differencesin silicon deposition. In barley, silicon was present in allthe walls of the endodermal cells, whereas in oats it was onlylocated in the inner tangential and radial walls. Wheat showedcultivar differences, no silicon was detected in Capelle Desprez,but it was present in the thickened endodermis of Little Jossand Hustler. In all the samples studied silicon was absent fromthe sub-epidermal sclerenchyma layer. The results are discussed in relation to the possible functionsof the endodermis and the signficance of silicification. Hordeum sativum Jess, barley, Avena sativa L, oat, Triticum aestivum L, wheat, silicon deposition, electron-probe microanalysis     

Pectin Depolymerase Activities Associated with Cell Walls from Cicer arietinum L. Epicotyl

The hydrolytic activity of the proteins extracted with 3 M LiClfrom chick-pea (Cicer arietinum) cell walls to pec-tic fractionsextracted with 50 mM trans-l,2-diaminocy-clohexane-N,N,N,N,-tetraaceticacid (CDTA) and 50 mM sodium carbonate was studied. The pecticfractions contained acidic polysaccharides with high molecularmass (higher than 5 x 10³ kDa), mainly composed of uronic acids,galac-tose, arabinose and rhamnose. The extracted proteins depo-lymerizedthe pectic polysaccharides and also a commercial preparationof polygalacturonic acid from citrus, detected by a decreasein their viscosity and a shift of their molecular mass distribution.The extract was able to depolymerize a uronic acid-rich componentin all the cases, although in different extent. Also, with regardto the CDTA-soluble pectins, a degradation of polyuronide anda shift of the molecular mass distribution of arabinogalactanwas observed. (Received March 11, 1997; Accepted September 10, 1997)     

Effects of Ascorbate on the Oxidation of Derivatives of Hydroxycinnamic Acid and the Mechanism of Oxidation of Sinapic Acid by Cell Wall-Bound Peroxidases

A cell wall fraction isolated from epicotyls of Vigna angularis,which contained both ionically and covalently bound peroxidases,rapidly oxidized p-coumaric, caffeic and ferulic acids and slowlyoxidized sinapic acid. The oxidation of sinapic acid was greatlyenhanced in the presence of p-coumaric, caffeic or ferulic acid.Ascorbate (20 µM) inhibited the oxidation of ferulic acidby about 70% and completely inhibited the oxidation of p-coumaricand ferulic acids. The cell wall fraction was capable of bindingferulic and sinapic acids but not caffeic acid. p-Coumaric acidbound only slightly to cell walls. The oxidation of p-coumaricand ferulic acids by KCl-washed cell walls was inhibited byabout 60% and 10%, respectively, by 20 µM ascorbate, butthe oxidation of caffeic acid was completely inhibited by ascorbateat less than 20 µM. The oxidation of derivatives of hydroxycinnamicacid by peroxidases released from cell walls by washing with1 M KCl was completely inhibited by ascorbate. These resultssuggest that the inhibition by ascorbate depends on the substituentgroup of the phenyl ring of the derivatives of hydroxycinnamicacid when the oxidation reaction is catalyzed by cell wall-boundperoxidases and that the oxidation of sinapic acid is mediatedby phenoxyl radicals of derivatives of hydroxycinnamic acidother than sinapic acid. (Received December 2, 1993; Accepted March 3, 1994)     

Autolysis Promotes the Extension Capacity of Zea mays Coleoptile Cell Walls in Response to Acid pH Solutions

The relationship between autolytic degradation of ß(1–3),(1–4)-D-glucanand acid pH-induced extension of isolated Zea mays cell wallshas been investigated using a constant-load extension technique.Acidic buffer (4.5) was able to induce an additional extension(E_a) on cell walls already extended at pH 6.8 buffer under a20 g-mass load, indicating that the additional extension (E_a)was the parameter that better represented the effect of thedifferent treatments on the mechanical properties of maize coleoptilecell walls. The additional extension in response to acidic pHwas higher when cell walls had been previously autolysed for24 h at pH 5.5. Furthermore, the acid-pH effect was dependenton the presence during the constant load extension of some thermo-labilefactors, suggesting the participation of expansins. Acid pHincreased E_a of native cell walls through an increase in theplastic extension (E_p) in agreement with a one step mechanismleading directly to irreversible (plastic) wall extension assuggested by Cosgrove (1977). The autolytic degradation of ß(1–3),(1–4)-D-glucan was also able to modify the mechanicalproperties of maize coleoptile cell walls increasing its elasticextension (E_e) in response to pH 4.5 buffer but that modificationonly leads to an increase in wall extension when expansins areactive, suggesting a cooperation between ß-glucanturnover and expansin action. (Received August 5, 1998; Accepted March 16, 1999)     

The Role of Cellulase in Hyalocyst Pore Formation in the Moss Syrrhopodon texanus

The sheathing leaf bases of Syrrhopodon texanus are primarilycomposed of porose, thin-walled cells called hyalocysts. Largepores develop in several of the hyalocyst walls through thegradual removal of wall material. A cellulase with a pH optimumof 4.5 was detected in extracts of Syrrhopodon gametophoresby viscometric assays and dinitrosalicylic acid assay for reducingsugars. Cytochemical localization of cellulase activity in associationwith thinning hyalocyst cell walls implicate this enzyme inpore formation and is the first direct evidence of cellulaseactivity in a member of the Bryophyta. Syrrhopodon texanus, cellulase, cytochemistry, Bryophyta, hyalocyst     

The Hypostase in Torenia fournieri Lind.: A Histochemical Study of the Cell Walls

A histochemical investigation on the cell walls of the hypostasein Torenia fournieri Lind. (Scrophulariaceae) revealed thatthey contain large amounts of callose, cellulose and pectins.Except in the middle lamellae, tests failed to show lignin inthe walls. It is surmised that the callose in the hypostasedevelops in order to regulate the flow of metabolites to theembryo sac. Torenia fournieri Lind., hypostase, cell wall, callose     

Chloroplast Doublets and Differential Staining of the Cell Walls in Photosynthetic Tissues of Amaranthus dubius Mart

Ultrastructural studies of the leaves of Amaranthus dubius revealpresence of chloroplast doublets in mature leaf tissues. Theyalso highlight the difference in the structural organisationof the bundle sheath and the mesophyll cell walls, a featurewhich may have functional significance. Amaranthus dubius Mart., Calaloo, mesophyll, bundle sheath, chloroplast doublets, plastid fusion, plastid division, differential staining of the cell walls     

Phototactic Chloroplast Displacement in the Photosynthetic Mutant, Lemna paucicostata Strain 1073

The photosynthetic mutant, strain 1073, of Lemna paucicostataTorr. (L. perpusilla Hegelm.) which has a block in the electrontransport chain between plastoquinone and cytochrome f is capableof light-induced chloroplast displacement movements. At 8000–14000 lx, chloroplasts of the mutant move from their positionadjacent to the inner periclinal wall of the mesophyll cellsto the anticlinal walls, i.e. along those walls parallel tothe direction of the light. Light does not appear to enhancerespiration of the photosynthetic mutant or of the wild typestrain (6746). These and other results support the idea thatchloroplast displacement in light is not solely the result oflight effects on photosynthesis and respiration. Lemna paucicostata Torr., photosynthetic mutant, phototaxis, chloroplast displacement     

Sieve Elements of Minor Veins in the Leaves of Beta vulgaris L.

End walls of sieve elements of minor veins of the leaves ofBeta vulgaris L. do not contain the multi-perforate sieve plateswhich typically occur on the end walls of sieve-tube membersof major veins. Instead, both end and side walls of the sieveelements of minor veins contain scattered pores which may occursingly or in small numbers. These pores are similar to thosewhich are grouped in sieve plates of major veins in size, possessionof callose and plugs of filaments. In addition to these pores,there are tubular connections 0.1 µ in diameter throughcharacteristically thickened parts of the cell wall betweensieve cells and companion cells. Sieve elements of minor veinsdiffer from those of major veins in structure as well as infunction.     

The Development of the Fruit Walls in Carya

Both the pistillate flowers and the developing fruits of CaryaovataandC. cordiformis are covered with secretory and protectivehairs. There are two kinds of the secretory hairs. In the morefrequent one the volatile oils are accumulated under the cuticlecovering the cells of the head. In the other the secretory substancesare accumulated inside the cells. The flowers and the developingfruits of the two investigated species have quite large emergenceson which the stomata most often occur. During the developmentof the fruit lenticels are formed in the green hull. Tanninsare accumulated not only in the green hull, but also in thecarpellary layer and in the packing tissue. There are some differences between C. ovata and C. cordiformisin the course of sclerification of the green hull. In C. cordiformisthe hull is less sclerified than in C. ovata, and the sclerificationof the carpellary layer occurs later. At a certain period ratherlarge gaps are present in the lignifying shell (in the regionsof the sutures) where the cells have thin cellulose walls. Thesegaps are partly filled with sclereids. The gaps become reducedto narrow chinks, in which the cell walls remain unlignified.The hard shell is not formed in all its width at the same time.During the process of formation of the hard fruit shell theremay arise single sclereids surrounded by thin-walled cells.Within a short time these undergo a similar modification andthe tissue becomes uniform as sclerenchyma. The sclereids ofthe hard shell have, in general, strongly folded cell walls.Owing to this they overlap each other.