Water relations, xylem embolism and histological features of Pinus thunbergii inoculated with virulent or avirulent pine wood nematode, Bursaphelenchus xylophilus

The development process of pine wilt disease caused by Bursaphelenchusxylophilus (Steiner and Buhrer) Nickle, pine wood nematode,was studied ecophysiologically and histologically in relationto pathogenicity of B. xylophilus. Judging from the predawnxylem pressure potential of needles, the heat pulse velocity,and the soil water potential, the control Pinus thunbergii Parl.,used for the study, was not water-stressed. Virulent B. xylophilusisolate can kill non-waterstressed pines. In virulent B. xylophilusisolateinoculated pines, the predawn xylem pressure potentialof needles abruptly decreased when the colour of 1-year-oldneedles changed to brown and then the water conducting functionof the xylem was lost completely. Avirulent B. xylophilus isolatedoes not affect the needle colour and the xylem pressure potentialof pines. Avirulent B. xylophilus isolate-inoculated pines,however, responded to nematode invasion by decreasing hydraulicconductance of stem and root xylems. In addition, oleoresinexudation slightly decreased. The decreased hydraulic conductanceresults from embolism of tracheids caused by cavitation in thecentral part of the xylem. From histological observation, allof the parenchyma cells in virulent B. xylophilus isolateinoculatedpines died. In contrast, the parenchyma cells, degenerated inavirulent B. xylophilus isolateinoculated pines, were limitedin the embolized region of the xylem. The difference betweenthe response of pine to the virulent B. xylophilus isolate invasionand that to avirulent B. xylophilus isolate invasion indicatesthat nematode-induced death of pine relates to the death ofparenchyma cells, as well as the decrease in xylem hydraulicconductance. Key words: Embolism, hydraulic conductance, parenchyma cells, pathogenicity of pine wood nematode, pine wilt disease.     

DoesAcetobacter diazotrophicusLive and Move in the Xylem of Sugarcane Stems? Anatomical and Physiological Data

We have previously shown that the nitrogen-fixing endophyteof sugarcane,Acetobacter diazotrophicuslives in the sugar solutionin the intercellular-space apoplast of the stem cortex. Variousauthors have claimed that it inhabits the xylem apoplast. Thispossibility has been investigated in the clone Ja 60-5 and shownto be most unlikely for the following reasons: (1) an adequatecarbon source is lacking in the xylem sap, and cannot be suppliedfrom the intercellular-space apoplast of the cortex. Diffusionof solutes into and out of the vascular bundles is preventedby complete lignification and suberization of the bundle sheathcell walls except at the nodes. (2) Longitudinal movement ofparticles as large as bacteria is severely limited at the nodes.Vessel end walls were found in 90% of vessels at each node,and only 1% of open vessels extended through two nodes. Noneextended as far as three nodes. In addition to vessel end walls,vessel continuity at nodes was interrupted by living cells.Dye solution in the transpiration stream in metaxylem vesselsdid not pass through these living cells, but accumulated incrystals (sump formation) in the vessels below the node. Onlyin some protoxylem vessels and cavities did dye solution movethrough many nodes. It is likely that selection of sugarcaneclones such as Ja 60-5 for resistance to bacterial wilt diseaseshave selected for clones that have limited vessel continuity.(3) When culturedA. diazotrophicuswas introduced into the transpirationstream, the xylem parenchyma reacted by secreting a bright redpolymer which killed the bacteria and blocked the movement ofwater. We conclude that the xylem flow-apoplast of this cloneof sugarcane is an unsuitable habitat forA. diazotrophicusandthat additional habitats to those of the intercellular-spaceapoplast should be sought elsewhere. Acetobacter diazotrophicus; endophytic bacteria; nitrogen fixation; sugarcane; vessel end walls; xylem apoplast; xylem bacteria; xylem segmentation     

The Mechanical Properties of Xylem Tissue from Tobacco Plants (Nicotiana tabacum'Samsun')

Tobacco plants (Nicotiana tabacum‘Samsun’) havebeen grown with an antisense CAD (cinnamyl alcohol dehydrogenase)gene. This modifies lignin production, resulting in lignin witha greater aldehyde content which is easier to extract chemically.This lignin probably has a reduced crosslink density. The changedproperties of the lignin affect the longitudinal tensile modulusof the xylem tissue (wood), reducing it by one third, from 2.8GPa to 1.9 GPa. Tobacco xylem tissue cell walls are more sensitiveto changes in the properties of the matrix than can be predictedusing current cell wall mechanical models.Copyright 1998 Annalsof Botany Company Tobacco,Nicotiana tabacum, xylem tissue, Young's modulus, matrix polymer connectivity, plant biomechanics.     

Vascular Development and Sap Flow in Apple Pedicels

Xylem and phloem tissues of the pedicel of apple fruit increasein cross-sectional area throughout development. The increasein phloem is similar in the two cultivars examined (Cox's OrangePippin and Royal Gala) and reflects a steadily increasing phloemsap flow to the fruit. The increase in xylem tissue is due toa proliferation of non-conducting, structural, components sinceclose examination reveals no increase in the number of vesselelements from just after flowering onwards. The greater number,and the larger diameter, of the vessels in Cox's explains theinitially higher xylem conductance found in this cultivar. In vitro measurements of xylem exudation reveal a decline duringthe growing season in the xylem conductance of both cultivarsand an increasing proportion of fruit (particularly in Cox's)in which the xylem comes to be totally non-conducting. Thisobservation is in line with previously reported measurementsof xylem sap flow in vivo. The straightforward techniques used in this study offer a feasiblealternative to more arduous methods of assessing xylem and phloemsap flows and their balance during growth.Copyright 1994, 1999Academic Press Apple, xylem, phloem, vascular development, sap flow, Malus domestica Borkh     

The Growth Response of the Stems of Genetically Modified Tobacco Plants (Nicotiana tabacum'Samsun') to Flexural Stimulation

Genetically modified tobacco plants (Nicotiana tabacum‘Samsun’)with antisense cinnamyl alcohol dehydrogenase DNA, produce secondaryxylem of a reduced tensile stiffness. These plants were grownalongside control plants. The stems of the plants were flexedor protected from flexing over a period of several weeks. Thetensile moduli and second moments of areas of the differenttissues inside the stems were measured and used to calculatethe bending stiffness of the plants. In tobacco, the cylinderof xylem was found to be the most important tissue in determiningthe bending stiffness of the plants. The thickness of the xylemtissue cylinder increased when plants were subjected to flexuralstimulation. This increased the bending stiffness of the stems.The response to mechanical stimulation was found to be correlatedwith tissue strain and the genetically modified plants wereable to exactly compensate for the reduced modulus of theirxylem tissue by increasing the thickness of the xylem tissuecylinder more than in control plants.Copyright 1999 Annals ofBotany Company. Tobacco plants, stem bending, xylem tissue, second moment of area, thigmomorphogenesis, mechanical strain.     

A Histopathological Study of Anomalous Growth in the Stem of Coccinia indica W. & A. Infested with Neolasioptera cephalandrae Mani

Gall formation due to attack of the larvae of Neolasiopteracephalandrae is common on the stem of Coccinia indica. Isolatedpatches of secondary meristematic centres develop throughoutthe proliferated tissue and they often differentiate into vasculartissues (either xylem or phloem or both). The larvae bore throughthe stem and live in lysigenous cavities in the ground tissue.Proliferation of cells of the secondary meristematic centresresults in gall formation and the normal vascular pattern isdisturbed and vascular bundles crushed. Development of mechanicaltissues in the affected region is suppressed. Additional cambiallayers with no definite pattern of orientation also developin the ground tissue and these produce patches of xylem tissue.Larval cavities are irregular and are surrounded by a nutritivetissue rich in protein and polysaccharides.     

A critical re-examination of pressure-volume analysis of conifer shoots: comparison of three procedures for generating PV curves on shoots of Pinus resinosa Ait. seedlings

Prediction of water relations attributes for red pine (Pinusresinosa Ait.) derived from pressure-volume (PV) curves varieddepending on which of three methods was used. The sap expressionmethod entailed the enclosure of a shoot in a pressure chamberand expression of xylem sap by applying a constant selectedpressure until sap flow ceased, at which point xylem water potentialand shoot weight were measured. A sap expression PV curve wasformed by aggregating pairs of water potential-weight measurements,each pair supplied by one of 25 shoots. The repeat pressurizationmethod involved repeatedly measuring xylem water potential andshoot weight on a single shoot drying on a laboratory bench.Repeat pressurization PV curves were constructed from data providedby a single shoot. The composite method utilized single measurementsof xylem water potential and shoot weight on 25-30 differentshoots ranging in relative water content from about 1.0 to 0.5achieved by bench drying. Composite PV curves were constructedfrom aggregate data supplied by a population of shoots. Therewas close agreement in all PV attributes generated using repeatpressurization and sap expression methods. In contrast, withthe composite PV method, there was a fundamental differencein the slope of the linear region of the PV curves, causingosmotic potentials at full turgor and turgor loss to be morenegative, and relative water content at turgor loss to be lowerand symplast fraction to be higher. Comparison of compositeand repeat pressurization PV curves over the same ranges inwater content did not eliminate differences in derived waterrelations attributes. Differences in water potential isothermsrelated to the PV procedures used suggest that prolongedor repeatedexposure to gas at high pressure may introduce errors in theestimation of water relations attributes. Key words: Pinus resinosa, pressure chamber, pressure volume, tissue water relations     

Seasonal Water Acquisition and Redistribution in the Australian Woody Phreatophyte, Banksia prionotes

Sap flows in the xylem of plant roots in response to gradientsin water potential, either between soil and atmosphere (transpiration)or soil layers of different moisture content (termed hydraulicredistribution). The latter has the potential to influence waterbudgets and species interactions, but we lack information forall but a few plant communities. We combined heat pulse measurementsof sap flow with dye and isotope tracing techniques to gaugethe movement of xylem sap within, and exudation from, rootsof Banksia prionotes (Lindley). We demonstrated ‘ hydrauliclift’ during the dry season and provide some evidencethat extremely dry soils limit hydraulic lift. In addition wereport difficulties posed by spiralled xylem tissue in rootsfor the application of heat pulse techniques. Copyright 2000Annals of Botany Company Banksia prionotes, sap flow, hydraulic lift, heat ratio method, deuterium, stable isotopes, root architecture.     

Haustorial Structure and Functioning of the Root Hemiparastic Tree Nuytsia floribunda(Labill.) R.Br. and Water Relationships with its Hosts

Observations on the origin and mature structure of the haustoriumof the Western Australian Christmas tree (Nuytsia floribunda)corroborate and extend the findings of earlier workers. We showthat the previously described sclerenchymatous ‘horn’or ‘prong’ formed within the haustorium acts asa sickle-like cutting device which transversely severs the hostroot and then becomes lodged in haustorial collar tissue directlyopposite to that where it originated. The cutting process isdeduced to be rapid and the gland-like fluid filled structurein the haustorium is suggested to generate a hydrostatic forcedriving the device through the host root. The haustorial parenchymacells at the tight junction between the endophytic part of thehaustorium and the cut face of the host root develop balloon-likeoutgrowths which intrude into the lumina of severed xylem vesselsof the host. Experiments feeding 0.05% (w/v) basic fuchsin tofreshly cut ends of host root segments distal to terminally-attachedmature haustoria demonstrate an apoplastic pathway from hostxylem elements fractured at the interface into haustorial parenchyma,and thence through vascular tissue to the haustorium into thetranspiring plant of Nuytsia. Application of labelled water(D₂O) to uncut basal roots of potted plants ofAcacia acuminataparasitized by Nuytsia results in labelling of leafy shootsof parasite and host, indicative of haustorial uptake of waterby Nuytsia from host root xylem in the intact association. Measurementsof xylem water potentials of pot-cultured seedling Nuytsia associatedwith a range of hosts, or of mature trees of Nuytsia and partnerwoody hosts in the native habitat, demonstrate consistentlymore negative potentials in the parasite than host, suggestingthat the parasite may regularly obtain xylem water through itshaustorial apparatus. Copyright 2000 Annals of Botany Company Root hemiparasite, Nuytsia floribunda, Loranthaceae, haustorial structure, host–parasite water relations     

Trans-Root Electrical Potential in Roots of Plantago media L. as Affected by Hydrostatic Pressure: The Induction of an O2 Deficient Root Core

Excised roots of Plantago media L. were subjected to small hydrostaticpressure gradients. Pressure applied with a displacement pumpeasily extracted gas present in cortical lacunae consequenton the gas saturation deficit of the bathing solution. The temporarypressure treatment resulted in an irreversible decrease in theO₂ diffusion rate through the root tissue and consequently inO₂ deficient root cores. Therefore electrogenic pumps located at the symplast/xylem interface,i.e. in the centre of the root, were inhibited by pressure. The longitudinal distribution of the xylem pumps could be computed:in Plantago media they are located in the basal parts of theroot. The effect of pressure on the electrogenic xylem pumpsdemonstrated here should be taken into account when the processof ion transport from the root to the shoot is studied whilethe root is pressurized. (Received December 5, 1983; Accepted March 26, 1984)     

On the Conductivity of the Xylem in Trees

The xylem of three tree species, the ring porous Fraxinus excelsiorL., and the diffuse porous Acer pseudoplatanus L., and Salixatrocinerea Brot., has been examined to determine how closelyflow through it obeys the Poiseuille equation. The equationwas most closly obeyed in the relationship between flow-rateand pressure difference between the ends of the xylem cylinder,though there were differences between the ring and diffuse poroustypes. The relationship of flow-rate and specific conductivityto the radius of the xylem cylinder was complex.Fraxinus showeda rise with increase of radius up to a given size (0.54cm),followed by a steep decline. This decline was shown to be causedby a progressive increase in the number of vessels which wereunable to conduct water. Small increases in the viscosity ofthe xylem liquid produced a disproportionately large fall inthe rate of flow, and the specific conductivity of the xylemdecreased. With viscosities greater than 2.5 centipoise, thespecific conductivity remained constant. An effect of alternatelyraising and lowering the hydrostatic pressure of the xylem wateron the specific conductivity of the wood is described. These results are discussed in relation to the structure ofthe xylem, and particularly with regard to the possibility thatthe conduction channels in the wood consist of a system of microcapillariessurrounding the vessel cavities.     

Modelling of the Partitioning, Assimilation and Storage of Nitrate within Root and Shoot Organs of Castor Bean (Ricinus communis L.)

An experimentally-based modelling technique was developed todescribe quantitatively the uptake, flow, storage and utilizationof NO₃-N over a 9 d period in mid-vegetative growth of sandcultured castor bean (Ricinus communis L.) fed 12 mol m^–3nitrate and exposed to a mean salinity stress of 128 mol m^–3NaCl. Model construction used information on increments or lossesof NO₃-N or total reduced N in plant parts over the study periodand concentration data for NO₃-N and reduced (amino acid) Nin phloem sap and pressure-induced xylem exudates obtained fromstem, petiole and leaf lamina tissue at various levels up ashoot. The resulting models indicated that the bulk (87%) of incomingnitrate was reduced, 51% of this in the root, the remainderprincipally in the laminae of leaves. The shoot was 60% autotrophicfor N through its own nitrate assimilation, but was oversuppliedwith surplus reduced N generated by the root and fed to theshoot through the xylem. The equivalent of over half (53%) ofthis N returned to the root as phloem translocate and, mostly,then cycled back to the shoot via xylem. Nitrate comprised almosthalf of the N of most xylem samples, but less than 1% of phloemsap N. Laminae of leaves of different age varied greatly inN balance. The fully grown lower three leaves generated a surplusof reduced N by nitrate assimilation and this, accompanied byreduced N cycling by xylem to phloem exchange, was exportedfrom the leaf. Leaf 4 was gauged to be just self-sufficientin terms of nitrate reduction, while also cycling reduced N.The three upper leaves (5–7) met their N balance to varyingextents by xylem import, phloem import (leaves 6 and 7 only)and assimilation of nitrate. Petioles and stem tissue generallyshowed low reductase activities, but obtained most of theirN by abstraction from xylem and phloem streams. The models predictedthat nodal tissue of lower parts of the stem abstracted reducedN from the departing leaf traces and transferred this, but notnitrate, to xylem streams passing further up the shoot. As aresult, xylem sap was predicted to become more concentratedin N as it passed up the shoot, and to decrease the ratio ofNO₃-N to reduced N from 0·45 to 0·21 from thebase to the top of the shoot. These changes were reflected inthe measured N values for pressure-induced xylem exudates fromdifferent sites on the shoot. Transfer cells, observed in thexylem of leaf traces exiting from nodal tissue, were suggestedto be involved in the abstraction process. Key words: Ricinus communis, nitrogen, nitrate, nitrate reduction, partitioning, phloem, xylem, flow models     

Does ABA in the Xylem Control the Rate of Leaf Growth in Soil-Dried Maize and Sunflower Plants?

Maize (Zea mays L.) and sunflower (Helianthus annuus L.) plantswere grown in large volumes of soil and leaf growth rate wasmonitored on a daily basis. Half the plants were given a soildrying treatment and when they showed a significant restrictionof growth rate (compared to both their daily growth rate beforedrying and the average growth rate of well-watered plants onthe same day), leaf water relations were measured and xylemsap was extracted using several techniques. There was a significant negative log-linear relationship betweenthe rate of leaf growth and the concentration of ABA in thexylem for both species. There was no clear relationship betweenleaf growth rate and leaf water potential or turgor for eitherspecies. Assessment of different methods for sampling xylemsap suggests that exudates collected from stem stumps or samplescollected by pressurizing the whole root system are suitablefor estimating ABA concentration in xylem, at least with largeplants of maize or sunflower, provided the first few hundredcubic millimetres of collected sap are used for the assay. Centrifugationof sections of stems resulted in dilution of ABA in the xylemsap with sap squeezed from parenchyma tissue. This is because,at least in plants subjected to mild soil drying, the concentrationof the ABA in the xylem is far higher than that in the cellsap of stem tissue. Results support the proposal that ABA plays a major role asa chemical signal involved in the root-to-shoot communicationof the effects of soil drying. The non-hydraulic restrictionof leaf growth by a chemical signal can be explained by theextra root-sourced ABA in the xylem and may be an importantcomponent of the modification of growth and development whichresults from prolonged soil drought. Key words: Soil drying, ABA, leaf growth, Zea mays L., Helianthus annuus L.     

Fusarium Wilt of Bananas in Jamaica: II. Some Apsects of Host-parasite Relationships

The incidence of Fusarium wilt in a given variety of bananais determined by numerous interacting factors such as geneticallydetermined resistance, physical and chemical soil conditions,and abundance of Fusarium oxysporum f. cubense. To attain markedresistance to wilt, ‘Gros Michel’ apparently requiresa combination of conditions; thus the soil must be alkaline,of medium to heavy texture, and rich in available potash. Nitrogenfertilizing tends to increase susceptibility to wilt. Some striking effects of weather on wilt incidence are describedand various aspects of crop management are discussed: soil deteriorationunder bananas undoubtedly contributed to development of wiltin the past. In the earliest stages of rootlet infection, the pathogen growsmore slowly in the resistant ‘Cavendish’ than inthe susceptible ‘Gros Michel’, probably owing tomore effective tissue reactions in the former. In the field,infections progressing along the root are often checked at theroot base: with ‘Gros Michel’, however, a smallproportion enter the rhizome and as a result wilt generallydevelops. Ability to check such infections, leading to recovery,is only exhibited on very fertile soils. The possible mode of action of various factors on wilt incidenceis discussed, and the importance of vigorous root growth inpromoting host resistance is emphasized.     

Three Different Methods for Measuring Xylem Cavitation and Embolism: A Comparison

Three different methods for measuring xylem embolism due towater cavitation were compared—the acoustic method, thehydraulic method and the anatomical method. Young plants ofCeratonia siliqua L. were water stressed for 9, 16 and 23 d. Xylem cavitation was detected by counting ultrasound (100–300kHz) acoustic emissions (AE) from 1-year-old twigs (acousticmethod). Xylem embolism was detected by measuring the loss ofhydraulic conductivity of twigs of the same age (hydraulic method).The blockage of single xylem conduits was detected by perfusingSafranin into the xylem of 1-year-old twigs of stressed plantsand measuring the number and the diameters of non-conductingxylem conduits, under the microscope (anatomical method). It was noted that: (a) the number of AE and the loss of conductivityincreased with the water stress applied; (b) a linear relationseemed to exist between the number of AE and the loss of conductivity,suggesting that the AE counted could be only (or mainly) producedin the xylem conduits; (c) the vulnerability of the xylem conduitsto embolism was a direct function of their diameter; and (d)the measured loss of conductivity was of the same order of magnitudeas the theoretical one. The three methods gave fairly similar results. Nonetheless,they are not alternative to one another in that: (a) the acousticmethod allows continuous recordings to be made but does notprovide information about the actual damage suffered by plants;(b) the hydraulic method is very informative but destructive;and (c) the anatomical method is very useful both in phytogcographicaland in genetic improvement studies. Ceratonia siliqua L., Carob tree, water stress, xylem embolism, acoustic method, hydraulic method, anatomical method     

Three Different Methods for Measuring Xylem Cavitation and Embolism: A Comparison

Three different methods for measuring xylem embolism due towater cavitation were compared—the acoustic method, thehydraulic method and the anatomical method. Young plants ofCeratonia siliqua L. were water stressed for 9, 16 and 23 d. Xylem cavitation was detected by counting ultrasound (100–300kHz) acoustic emissions (AE) from 1-year-old twigs (acousticmethod). Xylem embolism was detected by measuring the loss ofhydraulic conductivity of twigs of the same age (hydraulic method).The blockage of single xylem conduits was detected by perfusingSafranin into the xylem of 1-year-old twigs of stressed plantsand measuring the number and the diameters of non-conductingxylem conduits, under the microscope (anatomical method). It was noted that: (a) the number of AE and the loss of conductivityincreased with the water stress applied; (b) a linear relationseemed to exist between the number of AE and the loss of conductivity,suggesting that the AE counted could be only (or mainly) producedin the xylem conduits; (c) the vulnerability of the xylem conduitsto embolism was a direct function of their diameter; and (d)the measured loss of conductivity was of the same order of magnitudeas the theoretical one. The three methods gave fairly similar results. Nonetheless,they are not alternative to one another in that: (a) the acousticmethod allows continuous recordings to be made but does notprovide information about the actual damage suffered by plants;(b) the hydraulic method is very informative but destructive;and (c) the anatomical method is very useful both in phytogeographicaland in genetic improvement studies. Ceratonia siliqua L, Carob tree, water stress, xylem embolism, acoustic method, hydraulic method, anatomical method     

A New Theory for the Ascent of Sap--Cohesion Supported by Tissue Pressure

Recent work contradicting both the assumptions of the CohesionTheory, and the tensions measured in the xylem sap by the pressure-chamber,is reviewed. Measurements with the xylem-pressure probe revealpressures in vessels around 0 bar absolute, and no detectablegradients of pressure with tree height. Under high water stress,pressures down to -6 bar were found, but then cavitations occurredvery readily. Also, measurements of the cavitation thresholdsof water show an average threshold of about -2 bar. The uncertainfoundations of the Cohesion Theory are recalled from the yearsbefore 1965. Soon after that date, Scholander's measurementswith the pressure chamber were agreed to have confirmed thetheory and the existence of high tensions in the xylem. Before1965, many experiments over many years pointed to the conclusionsnow rediscovered, viz., no high tensions, and no gradients oftension. A resolution of these paradoxes is offered in the formof a new theory. This proposes that the driving force and thetransmission of the force are the same as in the Cohesion Theory,but the operating pressure of the xylem is raised into a stablerange by compensating tissue pressures pressing upon the trachearyelements. The tissue pressure does not propel the transpirationstream, which is still driven by evaporation, but protects thestream from cavitation. Evidence is presented for the existenceof positive pressures in roots, wood, and leaves. It is shownthat the anatomy of roots, wood, and monocotyledon and cryptogamvascular bundles is organized so that pressure is confined bymechanical barriers, and exerted upon the tracheary elementsby the living cells of the phloem and the xylem parenchyma.The Compensating-Pressure Theory also explains, among otherthings, root pressure, the function of the endodermis, the structureof wood, the constant association of xylem and phloem, the absenceof gas spaces in vascular tissue, the absence of a gravitationalgradient in the xylem, bleeding from cut palm inflorescences,how insects are able to withdraw sap from the xylem, and thevariable that is measured by the pressure chamber. This instrumentmeasures the water potential, but this is the potential notof xylem in tension, but of the compensating pressure appliedto the xylem. The requirements of the Theory are explained,and a number of predictions are made which are open to experimentaltesting.Copyright 1995, 1999 Academic Press Ascent of sap, cavitation, cohesion theory, endodermis, pressure chamber, root pressure, stem pressure, tissue pressure, transpiration, water potential, wood anatomy, xylem pressure     

The Development of the Runner-bean Leaf with Special Reference to the Relation between the Sizes of the Lamina and of the Petiolar Xylem: I. The Relation between Lamina Area and Petiolar Xylem

An investigation has been made of the relation between the sizesof the lamina and of the petiolar xylem of both mature and immatureleaves of the runner bean (Phaseolus multiflorus Willd.). The ratios xylem area/lamina area and the number of vessels/laminaarea are lowest for mature leaves. Immature leaves gave higherbut somewhat more variable values for these two ratios. There is a constant growth ratio between the lpminn area andthe xylem area, such that k is approximately o·61 inthe allometry formula. The significance of the results is briefly considered from thepoint of view that the xylem area is related to the water requirementsof the leaf.     

Modulation of eIF5A1 expression alters xylem abundance in Arabidopsis thaliana

Eukaryotic translation initiation factor 5A (eIF5A) is thoughtto facilitate protein synthesis by participating in the nuclearexport of specific mRNAs. In Arabidopsis, there are three isoformsof eIF5A. One of them, AteIF5A1, has been shown to be expressedin vascular tissue, specifically developing vessel members,using GUS as a reporter. In order to determine whether AteIF5A1plays a role in xylem formation, its full-length cDNA was constitutivelyover-expressed in transgenic Arabidopsis plants. Microscopicanalysis revealed that the cross-sectional area of the xylemin the main inflorescence stems of transgenic plants was 1.9-foldhigher than those of corresponding inflorescence stems of wild-typeplants. In wild-type stems, the primary xylem typically comprisedsix cell layers and was 105 µm thick, but increased to9–11 cell layers, 140–155 µm thick, in transgenicstems. Similarly, the secondary xylem increased from six celllayers, 70 µm thick, in control stems to 9 cell layers,95–105 µm thick, in transgenic stems. Moreover,constitutive down-regulation of AteIF5A1 using antisense technologyresulted in the major suppression of xylem formation comparedwith control plants, and the antisense transgenic plants werealso stunted. These data collectively indicate that eIF5A1 playsa role in xylogenesis. Key words: Arabidopsis thaliana, eukaryotic translation initiation factor 5A, inflorescence stem, xylem Received 5 November 2007; Revised 26 December 2007 Accepted 10 January 2008     

Water Pathways in Higher Plants: I. FREE SPACE IN WHEAT LEAVES

A technique has been developed for the study of pathways ofwater movement in the xylem and free space of wheat leaves.Plants were treated with Lead-EDTA chelate through either theroots or the leaves; after treatment the lead was precipitatedin situ as lead sulphide with hydrogen sulphide gas and itslocation determined by light and electron microscopy. Bulk water movement was in the lumen of the xylem, where therewas always a heavy deposit of lead sulphide after root treatments.Outside the xylem the deposits were confined to the cell wallsand were most dense in the middle lamella. Deposits were notfound in the cells themselves. The main zones of water loss,marked by heavy deposits of lead sulphide, were associated withthe stomata, the junctions of the periclinal walls of the epidermalcells, and the cuticle, leaf hairs, and specialized epidermalcells with pitted walls associated with the vascular bundles. Entry of lead chelate into the leaves was adequately describedby a diffusion model. The free space seemed to be located mainlyin the water of hydration of the pectin middle lamella and wasesmated to occupy 3 to 5 per cent of the volume of the tissue.