Differential allocation of constitutive and induced chemical defenses in pine tree juveniles: a test of the optimal defense theory

Optimal defense theory (ODT) predicts that the within-plant quantitative allocation of defenses is not random, but driven by the potential relative contribution of particular plant tissues to overall fitness. These predictions have been poorly tested on long-lived woody plants. We explored the allocation of constitutive and methyl-jasmonate (MJ) inducible chemical defenses in six half-sib families of Pinus radiata juveniles. Specifically, we studied the quantitative allocation of resin and polyphenolics (the two major secondary chemicals in pine trees) to tissues with contrasting fitness value (stem phloem, stem xylem and needles) across three parts of the plants (basal, middle and apical upper part), using nitrogen concentration as a proxy of tissue value. Concentration of nitrogen in the phloem, xylem and needles was found to be greater higher up the plant. As predicted by the ODT, the same pattern was found for the concentration of non-volatile resin in the stem. However, in leaf tissues the concentrations of both resin and total phenolics were greater towards the base of the plant. Two weeks after MJ application, the concentrations of nitrogen in the phloem, resin in the stem and total phenolics in the needles increased by roughly 25% compared with the control plants, inducibility was similar across all plant parts, and families differed in the inducibility of resin compounds in the stem. In contrast, no significant changes were observed either for phenolics in the stems, or for resin in the needles after MJ application. Concentration of resin in the phloem was double that in the xylem and MJ-inducible, with inducibility being greater towards the base of the stem. In contrast, resin in the xylem was not MJ-inducible and increased in concentration higher up the plant. The pattern of inducibility by MJ-signaling in juvenile P. radiata is tissue, chemical-defense and plant-part specific, and is genetically variable.     

Biomechanics of the columnar cactus Pachycereus pringlei

We report the longitudinal variations in stiffness and bulk density of tissue samples drawn from along the length of two Pachycereus pringlei plants measuring 3.69 and 5.9 m in height to determine how different tissues contribute to the mechanical stability of these massive vertical organs. Each of the two stems was cut into segments of uniform length and subsequently dissected to obtain and mechanically test portions of xylem strands, stem ribs, and a limited number of pith and cortex samples. In each case, morphometric measurements were taken to determine the geometric contribution each tissue likely made to the ability of whole stems to resist bending forces. The stiffness of each xylem strand increased basipetally toward the base of each plant where stiffness sharply decreased, reaching a magnitude comparable to that of strands 1 m beneath the stem apex. The xylem was anisotropic in behavior, i.e., its stiffness measured in the radial and in the tangential directions differed significantly. Despite the abrupt decrease in xylem strand stiffness at the stem base, the contribution made by this tissue to resist bending forces increased exponentially from the tip to the base of each plant due to the accumulation of wood. A basipetal increase in the stiffness of the pith (and, to limited extent, that of the cortex) was also observed. In contrast, the stiffness of stem rib tissues varied little as a function of stem length. These tissues were stiffer than the xylem in the corresponding portions of the stem along the upper two-fifths of the length of either plant. Tissue stiffness and bulk density were not significantly correlated within or across tissue types. However, a weak inverse relationship was observed for these properties in the case of the xylem and stem rib tissues. We present a simple formula that predicts when stem ribs rather than the xylem strands serve as the principal stiffening agents in stems. This formula successfully predicted the observed aspect ratio of the stem ribs (the average quotient of the radial and tangential dimensions of rib transections), and thus provided circumstantial evidence that the ribs are important for mechanical stability for the distal and younger regions of the stems examined.     

Localisation of plum pox virus in apricot stem and petiole tissues by tissue printing onto nitrocellulose membrane

The localisation of plum pox virus (PPV) in stem and petiole tissues of nine susceptible apricot cultivars and GF305 peach seedling has been studied. From stem and petioles consecutive transverse sections spaced at 1 mm were made and tissue sections printed onto nitrocellulose membrane. The resulting prints were probed with a specific antibody for plum pox virus, followed by a rabbit anti-goat antibody conjugated with horse radish peroxidase, in order to localise the virus within the tissues. In stems the virus was mainly present in xylem and pith. The possible presence of the virus in the sclerenchyma is discussed. In petioles the virus was present in epidermis and parenchymas, but not in vessels. The probable movement through the xylem and from cell to cell has been shown.     

Visualization of lateral water transport pathways in soybean by a time of flight-secondary ion mass spectrometry cryo-system

Water movement between cells in a plant body is the basic phenomenon of plant solute transport; however, it has not been well documented due to limitations in observational techniques. This paper reports a visualization technique to observe water movement among plant cells in different tissues using a time of flight-secondary ion mass spectrometry (Tof-SIMS) cryo-system. The specific purpose of this study is to examine the route of water supply from xylem to stem tissues. The maximum resolution of Tof-SIMS imaging was 1.8 μm (defined as the three pixel step length), which allowed detection of water movement at the cellular level. Deuterium-labelled water was found in xylem vessels in the stem 2.5 min after the uptake of labelled water by soybean plants. The water moved from the xylem to the phloem, cambium, and cortex tissues within 30-60 min after water absorption. Deuterium ion counts in the phloem complex were slightly higher than those in the cortex and cambium tissue seen in enlarged images of stem cell tissue during high transpiration. However, deuterium ion counts in the phloem were lower than those in the cambium at night with no evaporative demand. These results indicate that the stem tissues do not receive water directly from the xylem, but rather from the phloem, during high evaporative demand. In contrast, xylem water would be directly supplied to the growing sink during the night without evaporative demand.     

Ultrastructural study of the biodegradation processes. I. Beech (Fagus sylvaticus L.) leaf white rot

An electron microscopic study of beech leaf white rot shows a certain number of characteristic developmental stages which are identical whether the material is from in vitro experimentation or from natural incubation. Endowed with a cellulolytic properly seemingly localized in the apical region only, hyphae of the white rot agent only traverse the plant cell walls. Subsequently, hyphae penetrate condensed protoplasmic residues and make them progressively transparent to electrons. During this discoloring process, a lethal factor of as yet unknown nature appears, affecting other microorganisms already present in the leaves. Phloem and xylem vascular bundles do not present notable ultrastructural modifications. Therefore, leaf discoloration is not due to an alteration of the xylem constituents but to changes having occurred in the condensed cytoplasmic residues of the dead tissues.     

Woody plant and gravity]

In this review, we attempted to summarize the effect of gravity on growth of woody plants, broad leaved trees, on earth. It is well known that in tilted broad leaved trees, tension wood formed in the secondary xylem causes negative gravitropism. Gibberellin has been shown to induce tension wood in weeping branch, causing its upright growth. Recent study has shown that seedling of Japanese cherry tree grown on three dimensional clinostat, a device that simulates microgravity, grew at random angles, and that the formation of secondary xylem, as supporting tissue for upright growth, decreased. In the decreased xylem formation, the inhibition of the differentiation and development of fiber cell was clearly observed. These results suggest that in attitude control and morphogenesis of stem in woody plant, secondary xylem formation seriously relates to gravity on earth. In woody plant, the mechanism of gravity perception and the following signal transduction have not yet been elucidated, although the recent study reported the possibility that endodermal starch sheath cells and plant hormones may play some role in the mechanism. Space experiment for woody plant is expected to study these problem.     

Defense response of resistant host Impatiens balsamina to the parasitic angiosperm Cuscuta japonica

The response of the stem of a resistant host (Impatiens baslamina) to infection by the parasitic flowering plant Cuscuta japonica was studied with light and electron microscopy. The intra- and interfascicular cambial cells in the host stem first reacted to the penetrating upper haustorium by dividing, and the differentiation of the host xylem (vascular) tissues proceeded toward interfascicular areas from vascular bundles. When the host vascular tissue was invaded by the endophyte (haustorial portion in the host stem), the host xylem was displaced, and host vessels became occluded with parenchyma cells, resulting in tyloses. As the parasitism progressed, areas of the host stem penetrated by the endophyte became swollen via secondary growth and cell division in the parenchymatous cortex, pith, and interfascicular areas. During this intrusion by the endophyte, darkly stained necrotic reactions were detected at the interface between the host tissue and the invading endophyte. The results suggested that in the host tissues penetrated by the parasite, the formation of secondary tissue and swellings caused by active cell division of ground tissue and host vessel occlusion by tyloses constitute the host structural defense against the parasite.     

Influence of insulating dead leaves and low temperatures on water balance in an Andean giant rosette plant

Abstract. The influence of insulating dead leaves on water balance in Espeletia timotensis Cuatr., an Andean caulescent giant rosette plant, was studied under field and laboratory conditions. Removal of the dead leaf layer surrounding the stem changed the pattern of diurnal stem temperature variation and produced transient and permanent effects on water balance. The pattern of liquid water flow resistance increase at low stem temperatures suggested that much of the water flow in the stem was through living membranes, probably those of the pith cells. The pith was determined to be an important source of stored water for daily transpirational needs. The lethal effects of dead leaf removal were attributed to one or more of the following causes: (1) inhibition of pith recharge by subfreezing stem temperatures; (2) embolisms in stem xylem; (3) freezing injury to pith tissue. The results suggested that an insulating layer of marcescent leaves and the presence of an internal water reservoir closer to the rosette than the soil water are important adaptations for maintenance of a favourable water balance in tropical alpine habitats where freezing temperatures occur regularly but last only a few hours.     

Functional Imaging of Plants: A Nuclear Magnetic Resonance Study of a Cucumber Plant

Functional magnetic resonance imaging was used to study transients of biophysical parameters in a cucumber plant in response to environmental changes. Detailed flow imaging experiments showed the location of xylem and phloem in the stem and the response of the following flow characteristics to the imposed environmental changes: the total amount of water, the amount of stationary and flowing water, the linear velocity of the flowing water, and the volume flow. The total measured volume flow through the plant stem was in good agreement with the independently measured water uptake by the roots. A separate analysis of the flow characteristics for two vascular bundles revealed that changes in volume flow of the xylem sap were accounted for by a change in linear-flow velocities in the xylem vessels. Multiple-spin echo experiments revealed two water fractions for different tissues in the plant stem; the spin-spin relaxation time of the larger fraction of parenchyma tissue in the center of the stem and the vascular tissue was down by 17% in the period after cooling the roots of the plant. This could point to an increased water permeability of the tonoplast membrane of the observed cells in this period of quick recovery from severe water loss.     

A system for measuring penetration of radioactive ions across roots of intact plants

A technique was developed for estimating penetration of P³² across roots of intact plants (Phaseolus vulgaris) by measuring the level of isotope in the xylem stream. Penetration was defined as movement from the root surface to the xylem sap. The xylem sap measurement for P³² was made in the stem as the material ascended the plant in the transpiration stream. Stems were held near 0° to arrest metabolic concentration of isotope adjacent to the xylem column. A 3 layer environment control system was constructed to allow stem chilling in a manner that would not interfere with the environments of the roots or foliage. Despite these precautions, some extra-xylary build-up of ³²P occurred in the chilled stem. The mathematical function of the extra-xylary fraction was derived, and the difference between this value and total P³² in the stem represented xylem sap isotope.     

Myb genes from Hordeum vulgare: tissue-specific expression of chimeric Myb promoter/Gus genes in transgenic tobacco

The structures of the three Myb -related genes Hv1 , Hv5 and Hv33 from barley were determined. They contain a single intron located in the second repeat unit of the Myb -related domain. By analogy to the animal MYB oncoproteins this conserved region of the gene product was shown by filter-binding experiments to exhibit nucleic acid-binding activity. Tobacco plants transgenic for chimeric Myb promoter/ Gus genes express the enzyme in a developmentally controlled and tissue-specific manner. During germination and early stages of plant growth, GUS activity is seen in the root cap and adjacent meristematic tissue. At later stages of plant development, GUS activity is predominantly observed in the shoot apical meristem, the roots and the nodal regions of the stem. Within the stem at stages of secondary growth, Myb promoters are active in defined cell types. In the internode low GUS activity is displayed by the innermost cell layer of the cortex, the starch sheath, that surrounds the vascular cylinder of secondary xylem and phloem tissue, as well as in pith rays originating from vascular cambium initials. In the nodal region Myb promoter-controlled Gus expression is mainly confined to the abaxial starch sheath of the leaf trace, to the branch traces and to internal strands of primary phloem. It is suggested that in addition to their activity in meristematically active plant tissues Myb genes are expressed in conductive tissues that are closely associated with vascular bundles.     

Cytoskeletal organization during xylem cell differentiation

The water and mineral conductive tube, the xylem vessel and tracheid, is a highly conspicuous tissue due to its elaborately patterned secondary-wall deposition. One constituent of the xylem vessel and tracheid, the tracheary element, is an empty dead cell that develops secondary walls in the elaborate patterns. The wall pattern is appropriately regulated according to the developmental stage of the plant. The cytoskeleton is an essential component of this regulation. In fact, the cortical microtubule is well known to participate in patterned secondary cell wall formation. The dynamic rearrangement of the microtubules and actin filaments have also been recognized in the cultured cells differentiating into tracheary elements in vitro. There has recently been considerable progress in our understanding of the dynamics and regulation of cortical microtubules, and several plant microtubule associated proteins have been identified and characterized. The microtubules have been observed during tracheary element differentiation in living Arabidopsis thaliana cells. Based on this recently acquired information on the plant cytoskeleton and tracheary element differentiation, this review discusses the role of the cytoskeleton in secondary cell wall formation.     

Ion-mediated changes of xylem hydraulic resistance in planta: fact or fiction?

Although xylem provides an efficient transport pathway for water in plants, the hydraulic conductivity of xylem (K(h)) can still influence plant water status. For decades, the K(h) of functional xylem has been assumed to be constant in the short term because xylem consists of a network of dead interconnected capillary elements (conduits). Recent research has shown that K(h) can change in response to the cation content of the xylem fluid. Volume changes of pectin gel in nanometer-sized pores at inter-conduit connections are hypothesized to be the cause, and implications for xylem transport in planta are suggested. However, it seems too early to be conclusive about this phenomenon because the phenomenon has not been measured in planta with xylem fluids that realistically mimic natural xylem sap and the applied methods used to measure ion-mediated changes in K(h) have drawbacks.     

Pathways of infection of Brassica napus roots by Leptosphaeria maculans

Infection of Brassica napus cotyledons and leaves by germinating ascospores of Leptosphaeria maculans leads to production of leaf lesions followed by stem cankers (blackleg). Leptosphaeria maculans also causes root rot but the pathway of infection has not been described. An L. maculans isolate expressing green fluorescent protein (GFP) was applied to the petiole of B. napus plants. Hyphal growth was followed by fluorescence microscopy and by culturing of sections of plant tissue on growth media. Leptosphaeria maculans grew within stem and hypocotyl tissue during the vegetative stages of plant growth, and proliferated into the roots within xylem vessels at the onset of flowering. Hyphae grew in all tissues in the stem and hypocotyl, but were restricted mainly to xylem tissue in the root. Leptosphaeria maculans also infected intact roots when inoculum was applied directly to them and hyphae entered at sites of lateral root emergence. Hyphal entry may occur at other sites but the mechanism is uncertain as penetration structures were not observed. Infection of B. napus roots by L. maculans can occur via above- and below-ground sources of inoculum, but the relative importance of the infection pathways under field conditions is unknown.     

绞股蓝营养器官各发育阶段结构与总皂甙含量相关性的研究

应用组织化学定位技术及植物化学方法,研究了不同发育阶段绞股蓝茎叶的结构与其中人参皂甙分布积累和总皂甙含量的关系。绞股蓝内的人参皂甙通过特定的显色剂能显示从紫红到浅红的颜色反应,颜色深浅与人参皂甙含量呈正相关。结果表明,叶内的皂甙主要分布在叶肉组织内,未分化的叶肉组织比成熟组织含量高,衰老的叶片不含皂甙。茎内的皂甙主要分布于维管束的外生韧皮部薄壁细胞中,表皮、同化组织、厚角组织及周皮的栓内层也有少量分布,木质部和髓薄壁组织无皂甙积累;幼茎所含总皂甙高于发育成熟的茎和老茎,含量最高的则是具有贮藏功能的根状茎。在绞股蓝全草中,叶的总皂甙含量高于相应发育阶段的茎,而以当年生根状茎的含量最高。提出了皂甙在叶内积累、茎内运输和地下茎内贮藏的观点及科学采收绞股蓝的建议。     

The apoplastic continuum,nutrient absorption and plasmatubules in the dwarf mistletoeKorthalsella lindsayi (Viscaceae)

Summary The transfer of nutrients between host and parasite in mistletoes has generally been considered to occur via the xylem to xylem contacts at the host-parasite interface in the haustorial organ of attachment. A few workers, however, have recently begun to question this assumption and have suggested an alternative pathway of transport involving the intervening parenchyma cells which are often abundant in the parasite at the interface. But no morphological experimental evidence has yet been forthcoming in support of an apoplastic continuum across this interface between parasite and host.Our observations on the dwarf mistletoeKorthalsella lindsayi first indicate an absence of plasmodesmata at the interface, with the conclusion that symplastic transport between the two plants is not involved. However, application of apoplastic markers, such as Calcofluor white and lanthanum and uranyl ions, to the stem of the host results in the transfer of these tracers across the interface and into the tissues of the parasite. This demonstrates the existence of an apoplastic continuum between the two plants, and a pathway that is probably used in the normal transfer of water and other nutrients from host to parasite.From the apoplastic continuum provided by the walls of the haustorial parenchyma tissue, nutrients are transferred to the symplast for eventual distribution to other parts of the plant. Evidence for the active uptake of substances from the apoplast by the protoplasts of the parenchyma cells is shown by the convoluted appearance of the plasmalemma and its differentiation often into plasmatubules.     

Significant Difference in Hydrogen Isotope Composition Between Xylem and Tissue Water in Populus Euphratica

Deuterium depletions between stem water and source water have been observed in coastal halophyte plants and in multiple species under greenhouse conditions. However, the location(s) of the isotope fractionation is not clear yet and it is uncertain whether deuterium fractionation appears in other natural environments. In this study, through two extensive field campaigns utilizing a common dryland riparian tree species Populus euphratica Oliv., we showed that no significant δ¹⁸O differences were found between water source and various plant components, in accord with previous studies. We also found that no deuterium fractionation occurred during P. euphratica water uptake by comparing the deuterium composition (δD) of groundwater and xylem sap. However, remarkable δD differences (up to 26.4‰) between xylem sap and twig water, root water and core water provided direct evidence that deuterium fractionation occurred between xylem sap and root or stem tissue water. This study indicates that deuterium fractionation could be a common phenomenon in drylands, which has important implications in plant water source identification, palaeoclimate reconstruction based on wood cellulose and evapotranspiration partitioning using δD of stem water.     

Altitudinal Variations of Ground Tissue and Xylem Tissue in Terminal Shoot of Woody Species: Implications for Treeline Formation

1. The terminal shoot (or current-year shoot), as one of the most active parts on a woody plant, is a basic unit determining plant height and is potentially influenced by a variety of environmental factors. It has been predicted that tissues amount and their allocation in plant stems may play a critical role in determining plant size in alpine regions. The primary structure in terminal shoots is a key to our understanding treeline formation. The existing theories on treeline formation, however, are still largely lacking of evidence at the species level, much less from anatomy for the terminal shoot.2. The primary structures within terminal shoot were measured quantitatively for 100 species from four elevation zones along the eastern slope of Gongga Mountain, southwestern China; one group was sampled from above the treeline. An allometric approach was employed to examine scaling relationships interspecifically, and a principal components analysis (PCA) was performed to test the relation among primary xylem, ground tissue, species growth form and altitude.3. The results showed that xylem tissue size was closely correlated with ground tissue size isometrically across species, while undergoing significant y- or/and x-intercept shift in response to altitudinal belts. Further, a conspicuous characteristic of terminal shoot was its allocation of contrasting tissues between primary xylem and ground tissues with increasing elevation. The result of the PCA showed correlations between anatomical variation, species growth form/height classes and environment.4. The current study presents a comparative assessment of the allocation of tissue in terminal shoot across phylogenically and ecologically diverse species, and analyzes tissue, function and climate associations with plant growth forms and height classes among species. The interspecific connection between primary xylem ratio and plant size along an elevation gradient suggests the importance of primary xylem in explaining the treeline formation.     

Allometric co-variation of xylem and stomata across diverse woody seedlings

Leaf stomatal density is known to co-vary with leaf vein density. However, the functional underpinning of this relation, and how it scales to whole-plant water transport anatomy, is still unresolved. We hypothesized that the balance of water exchange between the vapour phase (in stomata) and liquid phase (in vessels) depends on the consistent scaling between the summed stomatal areas and xylem cross-sectional areas, both at the whole-plant and single-leaf level. This predicted size co-variation should be driven by the co-variation of numbers of stomata and terminal vessels. We examined the relationships of stomatal traits and xylem anatomical traits from the entire plant to individual leaves across seedlings of 53 European woody angiosperm species. There was strong and convergent scaling between total stomatal area and stem xylem area per plant and between leaf total stomatal area and midvein xylem area per leaf across all the species, irrespective of variation in leaf habit, growth-form or relative growth rate. Moreover, strong scaling was found between stomatal number and terminal vessel number, whereas not in their respective average areas. Our findings have broad implications for integrating xylem architecture and stomatal distribution and deepen our understanding of the design rules of plants' water transport network.     

Effects of temperature and sink activity on the transport of 14C-labelled indol-3yl-acetic acid in the intact pea plant (Pisum sativum L.)

The velocity and intensity of basipetal transport of ¹⁴C-labelled indol-3yl-acetic acid (IAA) applied to the apical bud of the intact pea plant were influenced by the temperature to which the stem was exposed and were not influenced by changes in the temperature of the root system when this was controlled independently between 5°C and 35°C. The velocity of transport increased steadily with temperature to a maximum in excess of 35°C and then fell sharply with further increase in temperature. The Q₁₀ for velocity, determined from Arrhenius plots, was low (ca. 1.3). Transport intensity increased to a maximum at about 25°C (Q₁₀=2.2) and then declined gradually with further increase in temperature. It is suggested that transport velocity and transport intensity are controlled independently.The characteristics of auxin transport through the stem were not affected by removal of the root system, or by the withdrawl of root aeration. Labelled IAA did not pass a region of the stem cooled to about 1.0°C, or through a narrow zone of stem tissue killed by heat treatment. In the latter case the heat treatment was shown not to interfere with the upward transport of water in the xylem. Labelled IAA continued to move into, and to accumulate in, the tissues immediately above a cooled or heat-killed region of the stem. It was concluded that the long-distance basipetal transport of auxin through the stem of the intact plant is driven by the transporting cells themselves and is independent of the activity of sinks for the transported auxin.The fronts of the observed tracer profiles in the stem were closely fitted by error function diffusion analogue curves. However, diffusion of IAA alone could not account for the observed characteristics of the transport and it is suggested that the curvilinear fronts of the profiles resulted from a diffusive mixing of exogenous IAA (or IAA-carrier complexes) with endogenous IAA already in the transport pathway.Abbreviations IAA indol-3yl-acetic acid - IAAsp indol-3yl-acetyl aspartic acid - CFM methyl 2-chloro-9-hydroxyfluorene-9-carboxylate (morphactin) - TIBA 2,3,5-triiodobenzoic acid - ABA abscisic acid