Reserve Lipids in Stem Root Primordia of Poplar

Lipids are reserve substances in stem root primordia of poplarand occur as droplets approximately uniformly in all cells ofroot primordia. In ultrathin sections they appear as sphericalelectron-dense particles (0.5–1.4 µm diameter),filling up virtually all the available space in the cytoplasm.The presence of lipids in stem root primordia was demonstratedin several species of the genus Populus from the section Aigeirosand Tacamahaca, as well as in another representative of thefamily Salicaceae, viz. in the genus Salix.     

The Effect of Calcium Starvation on Assimilate Partitioning and Mineral Distribution of the Phloem

Populus plants were grown in a medium lacking calcium and exposedto ¹⁴CO₂. In contrast to plants in the complete nutrient medium,the percentage amount of ¹⁴C-assimilates increased in the leavesof calcium-deficient plants and decreased in the stem and theroots. When plants were grown without potassium or magnesiumno differences in the amount of ¹⁴C-label occurred in comparisonwith plants in the complete nutrient medium. Translocation wasrecorded by microautoradiography. It was observed that considerableamounts of labelled photoassimilates were unloaded from thephloem in the middle part of the stem in plants of the completenutrient medium. In contrast, during calcium starvation ¹⁴C-labelwas restricted to the phloem of the stem. In addition, the concentrationsof magnesium and phosphorus showed a remarkable increase instem sieve tubes of calcium-deficient plants. When sieve tubesof source leaves from Populus, barley and maize were comparedwith those of sink leaves, the latter showed higher calciumconcentrations. The results suggest that calcium is a necessaryfactor in the regulation of phloem translocation. Key words: Calcium deficiency, phloem translocation, sieve element loading and unloading, X-ray microanalysis     

The Infection Biology of Sphaerulina musiva: Clues to Understanding a Forest Pathogen

Trees in the genus Populus and their interspecific hybrids are used across North America for fiber production and as a potential source of biofuel. Plantations of these species are severely impacted by a fungal pathogen, Sphaerulina musiva, the cause of leaf spot and stem canker. An inoculation protocol that does not rely on stem wounding to achieve infection was recently developed. Using this protocol two experiments were conducted to examine infection biology and disease development in the S. musiva-Populus interaction. In the first experiment non-wounded stems of one moderately resistant clone (NM6) and one susceptible clone (NC11505) were inoculated and examined by scanning electron microscopy at six different times (6 h, 12 h, 24 h, 72 h, 1 week, and 3 weeks) post-inoculation. The images indicate that the pathogen appears to enter host tissue through small openings and lenticels and that there are no significant differences in the penetration rate between the moderately resistant (NM6) and susceptible (NC11505) clones at 12 h post-inoculation. In a second experiment a histological comparison of stem cankers for resistant clone DN74 and susceptible clone NC11505 were conducted at three time points (3 weeks, 5 weeks, and 7 weeks) post-inoculation. Distinct differences in disease development were apparent between the resistant and susceptible clones at each time point, with the susceptible clone exhibiting a weak and delayed defense response. These results suggest, that following penetration, the pathogen may be able to interfere with the defense response in the susceptible host.     

The Cellular Pathway of Short-distance Transfer of Photosynthates and Potassium in the Elongating Stem ofPhaseolus vulgarisL. A Physiological Assessment

Plasmolytic disruption of plasmodesmata interconnecting metaphloemsieve element-companion cell complexes with small and largephloem parenchyma cells in the elongating region of internode2 ofPhaseolus vulgarisL. seedlings did not affect accumulationof phloem-imported¹⁴C-photosynthates and⁸⁶rubidium. The membrane-impermeantdye, 5(6) carboxyfluorescein, loaded into leaf phloem as themembrane-permeant diacetate ester, was found not to move radiallyout of the importing sieve elements in the internode elongationregion. In contrast, the apoplasmic tracer, Calcuofluor White,rapidly moved laterally throughout all tissues of the elongationzone. Hexoses, sucrose and potassium were identified as themain osmotica in internode apoplasmic sap. Label asymmetry in[¹⁴C](fructosyl)sucrose was retained on accumulation by excisedstem segments. Uptake of [¹⁴C]sucrose and⁸⁶rubidium by stemsegments exhibited saturation kinetics. Sucrose uptake was inhibitedby the slowly penetrating sulphydryl reagent, para-chloromercuribenzenesulphonicacid.In vitrorates of sucrose uptake, at apoplasmic concentrations,corresponded to its predictedin vivorate of delivery to thestem ground tissues from mature sieve elements when respiratorylosses were assumed to be confined to the stem phloem. For potassium,the total delivery rate could be accounted for by itsin vitrorateof uptake. Overall, it was concluded that radial transport,in the elongation zone of internode 2 ofPhaseolus vulgarisL.seedlings, follows an apoplasmic route from mature sieve elementsto stem ground tissues.Copyright 1998 Annals of Botany Company PhaseoluLes vulgaris, apoplasm, elongating stem, French bean, photosynthates, potassium, radial transfer, symplasm.     

Water Use of Kiwifruit Vines and Apple Trees by the Heat-Pulse Technique

Green, S. R. and Clothier, B. E. 1988. Water use of kiwifruitvines and apple trees by the heat-pulse technique.–J.exp. Bot. 39: 115–123. The compensation heat-pulse method has been used to measuresap velocities in the stem of kiwifruit vines (Actinidia deliciosa)and apple trees (Malus sylvestris x Red Delicious). Becauseof the high flow rates typical in kiwifruit vines, we were unableto measure heat-pulse velocity using standard probe spacings.We increased the spacing between sensors with the downstreamsensor 20 mm and the upstream sensor 5-0 mm respectively fromthe heater probe. Corrections for flow blockage by the probeswere re-calculated at this new spacing for our 2-0 mm-diameterheater and teflon temperature probes following the procedureof Swanson and Whitfield (1981) Sap flux through the stem was found from heat-pulse velocitiesat four radial depths in the stem. Fluxes measured using theheat-pulse technique were compared with water uptake from appletrees and kiwifruit vines that had been cut-off at the baseand the butt placed in a container of water. Heat-pulse measurementswere also compared with known flow rates through stem-sectionsof kiwifruit vine in the laboratory. In apple the heat-pulsemeasurements agreed with independent flux measurements in excisionexperiments. In kiwifruit the independently measured fluxeswere consistently 1.6 times larger than the fluxes measuredwith the heat-pulse method. Possible reasons for this anomalousresult in kiwifruit vines are discussed     

Vulnerability of xylem to embolism in relation to leaf water potential and stomatal conductance in Fagus sylvatica f. purpurea and Populus balsamifera

The vulnerability of xylem vessels to water stress-induced cavitationwas studied by measuring hydraulic conductivity and ultrasoundacoustic emissions (AEs) in Fagus sylvatica L. f. purpurea (Ait.)Schneid. and Populus balsamifera L. The occurrence of xylemembolism in summer was investigated in relation to leaf waterpotential and stomatal conductance. Populus was extremely vulnerableto cavitation, losing functional vessels due to embolism atwater potentials lower than –0.7 MPa. Fagus experiencedembolism when water potential fell below –1.9 MPa. Middaywater potentials often approached these threshold values. Whenevaporative demand increased rapidly on sunny days, water lossbecame limited by low stomatal conductance. Thus water potentialsfell only slightly below the threshold values inducing cavitation.Despite the differences in vulnerability, both species tolerateda similar embolism rate of about 10% in the summer. There wasno embolism reversal during prolonged periods of rain. AEs werepredictive of loss in hydraulic conductivity, indicating thatAEs were mainly confined to vessels. Finally, vessel lengthdistribution, vessel diameter (tangential axis), vessel density,and vessel wall thickness had been determined for both speciesinvestigated. Populus had longer and wider vessels than Fagus,whereas vessel wall thickness was similar in both species. Key words: Acoustic emissions, Fagus, Populus, stomatal closure, xylem embolism     

Action Potential in Luffa cylindlica and its Effects on Elongation Growth

The action potential of the non-seismonastic plant, Luffa cylindrica,was studied and characterized. Single action potentials of 30to 60 mV were evoked by electrical or cooling stimuli appliedto the stem internode. Action potentials were transmitted withoutdiminution along stimulated internodes shorter than 40 mm inlength. No difference in velocity between acropetal and basipetaltransmission was observed. This behavior is very similar tothat of Mimosa and Biophytum. The velocity of transmission ofaction potentials along internodes of Luffa was dependent upondistance from the point of stimulation; 10 to 20 mm sec^–1upto 40 mm, and 70 mm sec^–1 at around 50 mm from the pointof stimulation. The action potential was not transmitted tothe neighboring internode. Action potentials were also observedin the petiole and leaf. Beyond the threshold stimulus, theamplitude of the transmitted action potential depended on thestrength of the stimulus. The absolute and relative refractoryperiods for the transmission of action potentials were 2 minand 2 to 5 min, respectively. Elongation growth of the stem, recorded using a differentialtransformer, decreased after the generation of a single actionpotential in the upper most internode. This decreased growthrate continued for more than 5 min. (Received March 10, 1986; Accepted June 11, 1986)     

Does Cytokinin Transport from Root-To-Shoot in the Xylem Sap Regulate Leaf Responses to Root Hypoxia?

We have examined the hypothesis that cytokinins transportedfrom roots to shoots affects leaf growth, stomatal conductance,and cytokinin concentration of leaves of Phaseolus and a hybridpoplar (Populus trichocarpa x Populus deltoides) with hypoxicroots. Because cytokinins may interact with other substances,potassium and calcium concentrations were determined in xylemsap of Populus plants with hypoxic and aerated roots while gibberellin(GA) concentrations were measured in shoot tissues. Root hypoxiadecreased leaf growth and closed stomata in both species. Inboth species, fluxes of cytokinins out of the roots were reduced,but no differences in bulk leaf concentrations were measuredbetween the hypoxic and aerated plants. Shoots with aeratedroots contained slightly higher concentrations of GA₁ and GA₃than shoots from hypoxic plants. There were no differences incalcium or potassium concentrations in xylem sap between aerationtreatments. Exogenously applied cytokinins did not alleviatethe growth or stomatal responses caused by root hypoxia. Informationon the site(s) and mechanism(s) of cytokinin action and theways in which cytokinins are compartmentalized within plantcells will be required to understand the physiological significanceof cytokinin transport in the transpirational stream. Key words: Cytokinins, hypoxia, Populus, Phaseolus     

The Cellular Pathway of Short-distance Transfer of Photosynthates and Potassium in the Elongating Stem of Phaseolus vulgaris L. Stem Anatomy, Solute Transport and Pool Sizes

Based on an uniform elongation growth pattern and cellular structure,the apical 0·5-2·5 cm elongation zone of internode2 of Phaseolus vulgaris L. seedlings was selected as an experimentalsystem to study the radial pathway of photosynthate and potassiumtransfer from the phloem. An histological examination of thephloem within the elongation zone of internode 2 showed thatboth proto- and meta- phloem sieve elements were present. Theformer were fully differentiated at the commencement of elongationand became crushed as elongation proceeded. In contrast, about50% of the final number of metaphloem sieve element-companioncell complexes differentiated during the same period. The phloemdelivered some 99% of the sucrose and 72-82% of the potassiumaccumulated by the elongation zone. Solute budgets showed that,of the photosynthates and potassium entering the elongationzone, approximately 40% were retained and 60% transferred tothe shoot apex. Thus, the elongating stem acts not only as asignificant sink for photosynthates and potassium, but alsoas an axial phloem transport system to supply the shoot apex.Within the elongation zone, the principal tissue sinks weredetermined by the cellular localisation of [¹⁴C] photosynthatesand potassium by microautoradiography and ion electron microprobeanalysis respectively. About 80% of the photosynthates and potassiumwere located outside the phloem. The cortex and pith exhibitedthe greatest accumulation for photosynthates and the pith forpotassium.Copyright 1994, 1999 Academic Press Phaseolus vulgaris, elongating stem, French bean, photosynthates, potassium, radial transfer, stem anatomy, transport     

Effect of irradiance and vapour pressure deficit On stomatal response to CO2 enrichment of four tree species

The stomatal response of seedlings grown in 360 or 720 µmolmol^–1 to irradiance and leaf-to-air vapour pressure deficit(VPD) at both 360 and 720 µmol mol^–1 to CO₂ wasmeasured to determine how environmental factors interact withCO₂ enrichment to affect stomatal conductance. Seedlings offour species with different conductances and life histories,Cercis canadensis (L.), Quercus rubra (L.), Populus deltoides(Bartr. ex Marsh.) P. nigra (L.), and Pinus taeda (L.), weremeasured in hopes of identifying general responses. Conductanceof seedlings grown at 360 and 720 µmol mol^–1 CO₂were similar and responded in the same manner to measurementCO₂ concentration, irradiance and VPD. Conductance was lowerfor all species when measured at 720 than when measured at 360µmol mol^–1 CO₂ at both VPDs ({small tilde}1.5 and{small tilde}2.5 kPa) and all measured irradiances greater thanzero (100, 300, 600,>1600 µmol m^–2 S^–2)The average decrease in conductance due to measurement in elevatedCO₂ concentration was 32% for Cercis, 29% for Quercus, 26% forPopulus, and 11% for Pinus. For alt species, the absolute decreasein conductance due to measurement in CO₂ enrichment decreasedas irradiance decreased or VPD increased. The proportional decreasedue to measurement in CO₂ enrichment decreased in three of eightcases: from 0.46 to 0.10 in Populus and from 0.18 to 0.07 inPinus as irradiance decreased from>1600 to 100 µmolm^–2 s^–1 and from 0.35 to 0.24 in Cercis as VPD increasedfrom 1.3 to 2.6 kPa. Key words: Stomatal conductance, CO₂ enrichment, irradiance, vapour pressure deficit     

The role of apical development around the time of leaf initiation in determining leaf width at maturity in wheat seedlings (Triticum aestivum L.) with impeded roots

High soil resistance to root penetration (measured as penetrometerresistance, R_s slows down leaf growth and reduces mature leafsize in wheat seedlings {Triticum aestivum L.). Underlying changesin the kinetics of cell partitioning and expansion and in thesize and organization of mature cells were reported in companionpapers (Beemster and Masle, 1996; Beemster et al., 1996). Inthe present study, the relationships between apex growth, primordiuminitiation and expansion were analysed for plants grown at contrastingR_s, focusing on a leaf whose whole development proceeded afterthe onset of root impedance (leaf 5). High R_s reduced the rates of apex and leaf development, butdid not appear to have immediate effects on the pattern of developmentof the newly initiated phytomers. During an initial short period,the rate of development of a leaf primordium and associatednode were related to plastochronic age, according to similarrelationships (slopes) at the two R_s. Effects on developmentalpatterns were first detected on phytomer radial expansion duringplastochron 2. The ontogenetic pattern of leaf elongation wasaffected later, during the next few plastochrons preceding leafemergence (‘post-primordial stage’). It is concludedthat a reduction in the number of formative divisions and inthe number of proliferative cells along the intercalary mer-istemreported earlier (Beemster and Masle, 1996; Beemster et al.,1996) is not related to the size of the apical dome at leafinitiation nor to the size and number of meristematic cellsinitially recruited to the leaf primordium, which were all unaffectedby R_s. Rather they are generated at the primordial and post-primordialstages. Key words: Wheat, apex development, leaf primodium development, mature leaf width, root impedance     

Sucrose and Hexose Release by Excised Stem Segments of Vicia faba L.: The Sucrose-Specific Stimulating Influence of Cuscuta on Sugar Release and the Activity of Acid Invertase

By washing out solutes in 0.5 mM CaSO⁴at 25 °C during aperiod of 5–6 h, the release of sugars by excised stemsegments of Vicia faba L. was measured. The stem parasite Cuscutaeuropaea strongly stimulated the release of sucrose into theefflux medium; this effect was most marked during the last hoursof each experiment but this stimulating effect of the parasitecould not be detected for glucose and fructose. The fact thatparasitized stem segments released higher than normal hexoseamounts during the last hours of several experiments, couldbe explained as the result of extracellular hydrolysis of sucroseby free space invertase. A high free space acid invertase activitywas present in young stem segments of Vicia faba and in tissuesof Cuscuta. The stimulating influence of Cuscuta on sugar releaseby cells of stem segments appears sucrose-specific, supportingthe idea that the stimulating influence of Cuscuta on sugarrelease is restricted to the sieve-tube system. When metabolic inhibitors were added to the washing solutionor when segments were incubated at low temperature, no cleareffect of the parasite could be observed and for all segments(parasitized and non-parasitized) a strongly enhanced releaseof sucrose into the efflux medium was found during the lasthours of an experiment. These data support the idea that anintensive resorption of sucrose occurs within stem segments,after its release into the apoplast. Key words: Cuscuta europaea, Parasitic relationship, Phloem unloading     

Net CO2 Exchange Rate and Shoot Growth of Young Poplar (Populus) Clones

Eighteen poplar (Populus) clones, varying in growth capacitywere grown in plastic containers under outdoor conditions. Duringthe first year of growth their net CO₂ exchange rate (NCER)was studied by infra-red gas analysis as a function of photosyntheticphoton flux density (PPFD) under controlled environment conditions.Maximal NCER (under saturating PPFD) and the PPFD compensationpoint were significantly correlated with the first year's shootlength and hence with the above ground biomass production ofthe various clones. Key words: Poplar, Shoot growth, CO₂ exchange rate     

Loss of Stomatal Function in Ageing Hybrid Poplar Leaves

Under a variety of conditions old, non-senescent hybrid poplar(Populus sp.) leaves exhibited less stomatal control than young,mature leaves. Stomata of older leaves displayed oscillatorybehaviour more frequently, and oscillations were more random,than in younger leaves. Also, diffusive conductance of olderleaves changed less following sudden shifts from either darkto light, or vice versa, than in younger leaves, and temporalpatterns of diffusive conductance in older leaves appeared tobe relatively independent of microenvironmental conditions.Levels of conductance of older leaves were higher both in thedark and following excision than in younger leaves, while inthe light the situation was reversed. Total range of responseand stability of diffusive conductance were also lower in olderrather than in younger leaves. All of the observed age-relateddifferences suggest a loss of stomatal control with increasingleaf age. Leaf age, Populus sp., stomatal cycling, stomatal function, hybrid poplar     

Development of Stem Nodules in a Tropical Forage Legume, Aeschynomene afraspera

Rhizobial infection occurred on the stem of Aeschynomene afrasperaat the site of emergence of adventitious root primordia. Rhizobiainvaded cortical cells at the base of the root primordium. Thefirst infected cell enlarged and collapsed after rhizobia hadmultiplied in large numbers. At this time, a meristematic zonewas initiated some distance beneath the first infected cell.Rhizobial penetration into the deeper cortex was by progressivecollapse of infected cells towards the meristematic zone; rhizobiaentering the cortical cells by invagination of the host cellwall. At the entry point, rhizobia were embedded in digitatecell wall material. These infection structures were restricted,always originating from the cell wall of an adjacent infectedcell. Soon after infection, the cell collapsed progressivelyforming infection strand-like structures which developed upto the meristematic zone. When infection had reached the meristematiczone, invaded host cells ceased to collapse but divided repeatedlyto form the nodule. Key words: Aeschynomene afraspera, stem nodulation     

Cellular Structures, Plasma Membrane Surface Areas and Plasmodesmatal Frequencies of the Stem of Phaseolus vulgaris L. in Relation to Radial Photosynthate Transfer

At an early stage of secondary development, the metaphloem sieveelements appeared to be the only functional axial transportconduit in fully elongated stems of P. vulgaris plants. Thereis no apparent barrier to the radial transfer of solutes inthe stem apoplast. However, radial transfer through the stemsymplast could be limited by discontinuities resulting fromprotoplast degeneration of the protophloem fibres and developingsecondary xylem fibres. Estimates of possible sucrose fluxesthrough the apoplastic and symplastic routes indicated thatradial photosynthate transfer from the sieve element-companioncell (se-cc) complexes of the stem metaphloem could follow eithercellular route. In the case of apoplastic transfer, the plasmamembrane surface area of the se-cc complexes is only sufficientto support some form of facilitated movement of sucrose. Incontrast, the plasma membrane surface area of the phloem parenchymais sufficient to permit passive diffusion of sucrose to theapoplast. Plasmodesmatal frequencies suggest that any symplastictransfer to the phloem parenchyma from the sieve elements wouldbe via the companion cells. Phaseolus vulgaris, french bean, stem, photosynthate, radial transfer (photosynthates), cellular pathway     

The Penetration of Solar Radiation through Leaf Canopies of Different Structure

A comparative study has been made of the penetration of solarradiation through leaf canopies of contrasting structure. Forthis purpose experiments were undertaken on (a) Gladiolus withnearly vertical leaves, (b) Raphanus sativus with its rosettehabit, (c) two varieties of Linum usitatissimum, one an unbranchedflax type and the other a multi-stem linseed type, and (d) Agrostemmagithago where the apical leaves are sharply inclined and thebasal leaves disposed in a horizontal plane. Light gradientswere evaluated at different densities and at varying stagesof development by means of probes of silicon cells. Using afilter (cut off point 72-5 µ) simultaneous measurementswere made of the infra-red and ‘visible’ radiation.The distributions of the leaf and stem area through the canopywere recorded so that light interception per unit surface couldbe assessed. The shape of the light gradient between the top and bottom ofthe canopy was sigmoidal for Gladiolus but much less so forthe other species. Attenuation of the infra-red radiation withdepth was markedly smaller. For valid estimates of the extinction coefficient (K) it isessential to include stem areas; that is on theoretical groundshalf the total stem surface. The values of K were low for allspecies and in particular for Gladiolus. The relationship betweenlight attenuation and leaf area was approximately exponential.For Gladiolus it was established that K can decrease with increasingdensity but no changes with depth in the canopy were evident.The implications of these observations in terms of canopy structureare discussed.     

Techniques for enhanced release of leaf protoplasts in Populus

Microscopic examination of Populus leaf tissue following enzyme treatment revealed two factors contributing to low protoplast yields: (1) poor penetration of the enzymes into the tissue, and (2) entrapment of protoplasts in leaf debris during protoplast purification procedures. A simple combination of rapid grinding of the tissue in an Omni-mixer prior to enzyme treatment and forceful washing of leaf-debris after digestion provided high exposure of the cells, uniform digestion, and high yields of protoplasts of two Populus clones. Protoplasts exhibited cell wall regeneration and long-term viability in culture. The relative yield advantages of the techniques varied with the inherent digestibility of each clone but could produce up to 600 percent greater protoplast yields in a woody plant species in which protoplast isolation was previously limited. The techniques were also applicable to an herbaceous species, Solanum etuberosum.Abbreviations BA benzyladenine - NAA naphthalene acetic acid - WPM Woody Plant Medium, Lloyd and McCown (1980) - MS Murashige and Skoog Medium (1962) - (NC-XXXX) North Central Forest Experiment Station accession number assigned to Populus hybrid clones