Tracheary Elements Research Articles

Activation of ACS7 in Arabidopsis affects vascular development and demonstrates a link between ethylene synthesis and cambial activity.

Ethylene is a gaseous hormone that affects many processes of plant growth and development. During vascular development, ethylene positively regulates cambial cell division in parallel with tracheary element differentiation inhibitory factor (TDIF) peptide signaling. In this study, we identified an ethylene overproducing mutant, acs7-d, exhibiting enhanced cambial activity and reduced wall development in fiber cells. Using genetic analysis, we found that ethylene signaling is necessary for the phenotypes of enhanced cambial cell division as well as defects in stem elongation and fiber cell wall development. Further, the cambial cell proliferation phenotype of acs7-d depends on WOX4, indicating that the two parallel pathways, ethylene and TDIF signaling, converge at WOX4 in regulating cambium activity. Gene expression analysis showed that ethylene impedes fiber cell wall biosynthesis through a conserved hierarchical transcriptional regulation. These results advance our understanding of the molecular mechanisms of ethylene in regulating vascular meristem activity.

Variation in leaf anatomy, vascular traits and nanomechanical cell-wall properties among European beech (Fagus sylvatica L.) provenances

Structural and ultrastructural traits of leaf xylem vary among European beech ( Fagus sylvatica L.) provenances in relation to climate. High tracheary element lumen area was observed in central Europe and decreased towards the range margins, while tracheary element density displayed opposite trend. Leaf anatomy and xylem cell-wall deformation were associated with climate continentality at the sites of origin. Understanding structural and physiological properties of tree populations adapted to local climate in response to climate may be of direct relevance for forestry. Identifying patterns of heritable geographical variation and phenotypic associations of leaf-anatomical, vascular and cell-wall properties in European beech, a dominant tree species of European forests. Fifteen beech populations planted in a common-garden test were studied employing gas exchange and chlorophyll a fluorescence measurements as well as light and atomic force microscopies of leaf samples. Nanomechanical properties of leaf-xylem cell walls did not show a clear geographical trend, but were correlated with the temperature seasonality of the site of origin. Populations from eastern Germany, northern Poland and the Czech Republic showed lower tracheary element lumen area and higher tracheary element density compared with the other populations, while xylem conductivity negatively correlated with latitude. Thickness of leaf parenchyma layers increased towards the East and, consequently, correlated with temperature indicators of climate continentality. No correlation between structural and physiological traits was observed. The observed trends and associations with climate indicate adaptive variation of the studied traits.

Imperforate tracheary elements and vessels alleviate xylem tension under severe dehydration: insights from water release curves for excised twigs of three tree species.

PremiseWater stored in the xylem of woody plants is important for supporting the transpiration stream under prolonged drought, yet the source of stored water within the xylem during drought remains unclear. Insights into xylem water utilization during drought will uncover the adaptation strategies of the test species to stress.MethodsTo fill the existing knowledge gap, we excised twigs of Abies firma (Japanese fir, conifer), Cercidiphyllum japonicum (katsura tree, diffuse‐porous) and Quercus serrata (konara oak, ring‐porous) to quantify interspecific variation of water transfer in xylem corresponding with increasing cumulative water release (CWR) using micro x‐ray computed tomography and cryo‐SEM.ResultsFor all species studied, the main components of water storage within the operating range of water potential were not living cells but cavitation release and capillaries. Abies firma maintained water in the earlywood‐like cells, for possible maintenance of the transpiration stream. Cercidiphyllum japonicum maintained water in its vessels over 200 kg m‐3 of CWR, while Q. serrata lost most of its water in vessels with increasing CWR up to 100 kg m‐3. Cercidiphyllum japonicum exhibited a higher water storage capacity than Q. serrata. Under high CWR, narrow conduits stored xylem water in C. japonicum and imperforate tracheary elements in Q. serrata.ConclusionsAmong the species examined, increasing CWR appears to indicate differential utilization of stored water in relation to variation of xylem structure, thereby providing insight into the interspecific responses of tree species to drought.

Ecological wood anatomy of Alnus nepalensis (Betulaceae) throughout Nepal

Abstract Variation in the wood structure of Alnus nepalensis D. Don (Betulaceae) in relation to tree size and habitat altitude was studied in four areas of Nepal: Tamur, Rolwaling, Budi Gandaki, and Bajhang-Khaptad. Alnus nepalensis is a pioneer tree distributed in the Himalayas from northern India to southwestern China and grows throughout Nepal in a wide altitudinal range from 200 m in the tropical zone to 3030 m at the upper limit of the temperate zone. Ecological trends in the wood structure of Alnus nepalensis varied between the studied areas, distinct in the Tamur area, but indistinct in other areas, and tended to be detected in areas with specimens from altitudinal ranges of 2000 m. When all the specimens were separated at 1800 m in elevation and analyzed as two altitudinal groups, features of vessel size and distribution and the ratio of solitary vessels had a significant correlation with tree height in both groups, and tall trees tended to have larger, more solitary vessels. Vessel element and fiber lengths did not have any significant trends in the lower group but had a negative correlation with altitude in the upper group along with the reduction of tree height. Thus, vessel size seems to facilitate the height growth of this pioneer tree, and shorter tracheary elements and more vessel multiples seem to be related to the adaptation of smaller trees at higher elevations.

Discovering Biomass Structural Determinants Defining the Properties of Plant-Derived Renewable Carbon Fiber

SummaryTraditional lignocellulosic feedstock research has focused on biomass characteristics essential for improving saccharification efficiency, yet the key biomass features underlying high-quality renewable lignin materials remain unknown. Nevertheless, modern biorefinery cannot achieve sustainability and cost-effectiveness unless the lignin stream can be valorized. We hereby addressed these scientific gaps by investigating biomass characteristics defining lignin-based carbon materials properties. Lignin from eight sorghum samples with diverse characteristics was fabricated into carbon fibers (CFs). Remarkably, only lignin uniformity was found to define CF mechanical performance, highlighting the new structure-property relationship. Contrarily, lignin content and composition did not impact on carbon material properties. Mechanistic study by XRD and Raman spectroscopy revealed that higher lignin uniformity enhanced CF microstructures, in particular, turbostratic carbon content. The study for the first time highlighted lignin uniformity as an important biomass structure determinant for renewable products, which opened up new avenues for feedstock design toward diverse products enabling sustainable and cost-effective bioeconomy.

An auxin transport network underlies xylem bridge formation between the hemi-parasitic plant Phtheirospermum japonicum and host Arabidopsis.

Parasitic plants form vascular connections with host plants for efficient material transport. The haustorium is the responsible organ for host invasion and subsequent vascular connection. After invasion of host tissues, vascular meristem-like cells emerge in the central region of the haustorium, differentiate into tracheary elements and establish a connection, known as a xylem bridge, between parasite and host xylem systems. Despite the importance of this parasitic connection, the regulatory mechanisms of xylem bridge formation are unknown. Here, we show the role of auxin and auxin transporters during the process of xylem bridge formation using an Orobanchaceae hemiparasitic plant, Phtheirospermum japonicum The auxin response marker DR5 has a similar expression pattern to tracheary element differentiation genes in haustoria. Auxin transport inhibitors alter tracheary element differentiation in haustoria, but biosynthesis inhibitors do not, demonstrating the importance of auxin transport during xylem bridge formation. The expression patterns and subcellular localization of PIN family auxin efflux carriers and AUX1/LAX influx carriers correlate with DR5 expression patterns. The cooperative action of auxin transporters is therefore responsible for controlling xylem vessel connections between parasite and host.

Histopathological evidence of concomitant sexual and asexual reproduction of Elsinoë ampelina in grapevine under subtropical climate

Anthracnose caused by Elsinoë ampelina is one of the most serious diseases affecting grapevine production worldwide. Previous studies have described many structural and histochemical changes in grapevine leaves associated with E. ampelina colonization and the presence of ascus-like structures in the lesions. In this paper, we characterized the colonization and reproduction of E. ampelina in grapevine tendrils, stems and berries using light, epifluorescence and scanning electron microscopy. In the early stages of colonization, shape changes and the collapse of epidermal and parenchyma cells were more pronounced in the center of lesions. At this stage, pectic substances accumulated inside the parenchyma cells and hyphae were observed colonizing tissues inter- and intracellularly, including tracheary elements. As colonization evolved, lesions enlarged and infected tissues became necrotic and collapsed. Tyloses and conidiogenous cells producing conidia were observed in the tracheary elements. Conidiogenous cells and conidia were also present on the lesion surface. On the same lesions, bitunicate asci containing two-to three-celled ascospores were observed on the surface and throughout the stromatic tissue. Our analyses showed that both sexual and asexual stages of E. ampelina occur from the beginning of the epidemic, which may help to explain the explosive nature of the disease when early stages of grapevine development coincide with rainy seasons.

TDIF regulates auxin accumulation and modulates auxin sensitivity to enhance both adventitious root and lateral root formation in poplar trees.

Of six TRACHEARY ELEMENT DIFFERENTIATION INHIBITORY FACTOR (TDIF)-encoding genes in poplar, PtTDIF1 is predominantly expressed in adventitious roots (ARs), and the other five PtTDIFs are preferentially expressed in lateral roots (LRs). Upon auxin application, expression of all PtTDIFs declined in ARs but transiently increased in LRs. Both exogenous TDIF peptides and overexpression of PtTDIFs in poplar positively regulated the initiation and elongation of LRs, and overexpression of PtTDIFs also increased the number of ARs. As visualized by the auxin-responsive marker DR5:GUS, TDIF had differential impacts on the auxin signaling activity in ARs and LRs, which was corroborated by the free indole-3-acetic acid (IAA) measurements in them. Shoot tips of PtTDIF2- and PtTDIFL2-overexpressing (together as PtTDIFsOE) trees revealed an enhanced IAA biosynthetic capacity, and removal of the aerial tissues dramatically diminished the root phenotypes of micro-propagated PtTDIFsOE trees. Furthermore, PtTDIFsOE poplars displayed an increased sensitivity for exogenous IAA, and N-1-naphthylphthalamic acid (NPA) completely blocked the TDIF-induced AR and LR formation. In PtTDIFsOE roots, several auxin-related LR initiation markers such as GATA23, LBD16 and LBD29 were transcriptionally upregulated, further supporting that TDIF regulates LR organogenesis by strengthening the spatiotemporal auxin cues and that dynamic interplays between hormones govern root branching and developmental plasticity in tree species.

Active DNA demethylation regulates tracheary element differentiation in Arabidopsis.

DNA demethylation is important for the erasure of DNA methylation. The role of DNA demethylation in plant development remains poorly understood. Here, we found extensive DNA demethylation in the CHH context around pericentromeric regions and DNA demethylation in the CG, CHG, and CHH contexts at discrete genomic regions during ectopic xylem tracheary element (TE) differentiation. While loss of pericentromeric methylation occurs passively, DNA demethylation at a subset of regions relies on active DNA demethylation initiated by DNA glycosylases ROS1, DML2, and DML3. The ros1 and rdd mutations impair ectopic TE differentiation and xylem development in the young roots of Arabidopsis seedlings. Active DNA demethylation targets and regulates many genes for TE differentiation. The defect of xylem development in rdd is proposed to be caused by dysregulation of multiple genes. Our study identifies a role of active DNA demethylation in vascular development and reveals an epigenetic mechanism for TE differentiation.

Lignin synthesized by CmCAD2 and CmCAD3 in oriental melon (Cucumis melo L.) seedlings contributes to drought tolerance.

CmCAD2 and CmCAD3 function more positively than CmCAD1 in oriental melon for lignin synthesis which is important to ensure internal water status and thus for drought tolerance. Well-lignification may be the guarantee of efficient axial water transport and barrier of lateral water flow in oriental melon tolerating drought stress, however remains to be verified. As an important enzyme in monolignol synthesis pathway, five cinnamyl alcohol dehydrogenase (CAD) genes were generally induced in melon seedlings by drought. Here we further revealed the roles of CmCAD1, 2, and 3 in lignin synthesis and for drought tolerance. Results found that overexpressing CmCAD2 or 3 strongly recovered CAD activities, lignin synthesis and composition in Arabidopsis cadc cadd, whose lignin synthesis is disrupted, while CmCAD1 functioned modestly. In melon seedlings, silenced CmCAD2 and 3 individually or collectively decreased CAD activities and lignin depositions drastically, resulting in dwarfed phenotypes. Reduced lignin, mainly composed by guaiacyl units catalyzed by CmCAD3, is mainly due to the limited lignification in tracheary elements and development of Casparion strip. While CmCAD1 and 2 exhibited catalysis to p-coumaraldehyde and sinapaldehyde, respectively. Compared with CmCAD1, drought treatments revealed higher sensitivity of CmCAD2 and/or 3 silenced melon seedlings, accompanying with lower relative water contents, water potentials and relatively higher total soluble sugar contents. Slightly up-regulated expressions of aquaporin genes together with limited lignification might imply higher lateral water loss in stems of silenced lines. In Arabidopsis, CmCAD2 and 3 transgenic lines enhanced cadc cadd drought tolerance through recovering lignin synthesis and root development, accompanying with decreased electrolyte leakage ratios and increased RWCs, thus improved survival rates. Briefly, lignin synthesized by CmCAD2 and 3 functions importantly for drought tolerance in melon.

TDIF peptides regulate root growth by affecting auxin homeostasis and PINs expression in Arabidopsis thaliana.

TDIF and TDIF-like peptides in excess simultaneously facilitate primary root elongation and lateral root formation through regulating auxin distribution and transport. Tracheary element differentiation inhibitory factor (TDIF) plays key roles in mediating cell-cell communication and stem cell maintenance during vascular development. Recently, TDIF has also been linked to lateral root (LR) organogenesis through Brassinosteroid Insensitive 2 (BIN2) action. In this work, by comparing the in vitro and in vivo activities of AtCLE41-encoded TDIF and one poplar-derived TDIF-like peptide in Arabidopsis thaliana, we demonstrated that both TDIFs promoted primary root (PR) growth and stimulated LR formation. Without affecting auxin biosynthesis and catabolism, TDIFs suppressed the auxin maxima at PR apex but intensified the auxin accumulation at LR initiation sites along the longitudinal axis of PR. TDIF did not alter root sensitivity to exogenous auxin and mutants with varied endogenous auxin levels responded to TDIF peptides in a wild-type manner but to a lesser extent. Intriguingly, TDIF specifically upregulated the transcript abundance of PINs and multiple pin mutants displayed insensitivity to TDIF, demonstrating that PIN-mediated polar auxin transport (PAT) is indispensably required for the TDIF-induced root phenotypes. Taken together, our results revealed that TDIF might target PAT via mobilizing auxin efflux carriers to dynamically regulate the auxin signaling output and hence facilitate PR growth and LR formation.

Xylem vessel‐diameter–shoot‐length scaling: ecological significance of porosity types and other traits

AbstractFlowering plants predominantly conduct water in tubes known as vessels, with vessel diameter playing a crucial role in plant adaptation to climate and reactions to climate change. The importance of vessels makes it essential to understand how and why vessel diameter, plant height, and other ecological factors are interrelated. Although shoot length is by far the main driver of variation in mean vessel diameter across angiosperms, much remains to be understood regarding the factors accounting for the abundant variation around they‐axis in plots of mean species vessel diameter against shoot length. Here, we explore the potential role of porosity types, wood density, leaf phenology, background imperforate tracheary element type, vasicentric tracheids, vascular tracheids, perforation plate type, and successive cambia in causing variation in they‐intercept or slope of the mean species vessel‐diameter– and vessel‐density–shoot‐length associations at the shoot tip and base. We detected numerous cases of ecologically significant variation. For example, latewood vessels of ring porous species scale with a lower slope than earlywood, i.e., latewood vessels are relatively narrow in taller plants. This pattern is likely the result of selection favoring freezing‐induced embolism resistance via narrow vessels. Wood density was negatively associated with vessel diameter, with low wood density plants having wider vessels for a given height. Species with scalariform perforation plates scale with a lower shoot base vessel‐diameter–shoot‐length slope, likely reflecting selection against scalariform plates in wide vessels. In other cases, functional groups scaled similarly. For example, species with successive cambia did not differ from those with conventional single cambia in their mean vessel‐diameter–shoot‐length scaling, rejecting our prediction that species with successive cambia should have narrower vessels for a given shoot length. They did, however, have fewer vessels per unit shoot cross‐sectional area than plants of similar heights, likely because vessels have longer functional lifespans (and therefore are fewer) in species with successive cambia. Our methods illustrate how vessel diameter can be studied taking shoot length into account to detect ecologically important variation and construct theory regarding plant adaptation via the hydraulic system that includes plant size as a vital element.

WRKY15 Suppresses Tracheary Element Differentiation Upstream of VND7 During Xylem Formation.

Formation of the vascular cylinder, a structure critical to water and nutrient transport in higher plants, is highly regulated. Here we identify WRKY15 as an important regulator that suppresses tracheary element (TE) differentiation in Arabidopsis (Arabidopsis thaliana). Overexpression of WRKY15 resulted in discontinuous protoxylem vessel files and TEs with reduced spiral wall thickening/lignification. Expression of a dominant-negative WRKY15 variant, WRKY15-EAR, led to extra protoxylem vessels and ectopic TEs with increased spiral wall thickening/lignification. Ectopic TE formation in the root cortex and hypocotyl/leaf epidermis reveals that the suppression of WRKY15 is sufficient to trigger the transdifferentiation of other types of cells to TEs. Expression profiling, RT-qPCR, and reporter analyses revealed that WRKY15 suppresses the expression of VASCULAR-RELATED NAC DOMAIN7 (VND7), a master transcriptional regulator that promotes TE differentiation. We propose that WRKY15 negatively regulates VND7 expression indirectly based on (1) the absence of a W-box in the promoter of VND7 and (2) the observation that WRKY15 and VND7 are expressed in different cells in the vascular cylinder, with WRKY15 expressed in the procambial cells and VND7 in the protoxylem poles of procambium and differentiating TEs. Future research is needed to reveal the details underlying the interaction of WRKY15 and VND7 in plant vascular development.

Light affects tissue patterning of the hypocotyl in the shade-avoidance response.

Plants have evolved strategies to avoid shade and optimize the capture of sunlight. While some species are tolerant to shade, plants such as Arabidopsis thaliana are shade-intolerant and induce elongation of their hypocotyl to outcompete neighboring plants. We report the identification of a developmental module acting downstream of shade perception controlling vascular patterning. We show that Arabidopsis plants react to shade by increasing the number and types of water-conducting tracheary elements in the vascular cylinder to maintain vascular density constant. Mutations in genes affecting vascular patterning impair the production of additional xylem and also show defects in the shade-induced hypocotyl elongation response. Comparative analysis of the shade-induced transcriptomes revealed differences between wild type and vascular patterning mutants and it appears that the latter mutants fail to induce sets of genes encoding biosynthetic and cell wall modifying enzymes. Our results thus set the stage for a deeper understanding of how growth and patterning are coordinated in a dynamic environment.

Vegetative propagation of elite Eucalyptus clones as food source for honeybees (Apis mellifera); adventitious roots versus callus formation

Summer and autumn in Israel are highly arid with not enough plants in bloom offering nectar and pollen to support the local apiary. This leads to decline in colony health and honey production. To increase food sources for honeybees, we initiated a project to clone elite Eucalyptus trees exhibiting constant and rich blooming from late summer to early winter. We induced adventitious roots from cuttings of two mature Eucalyptus trees of which nectar production and honeybees’ attraction was measured: Eucalyptus brachyphylla and Eucalyptus x trabutii. During the rooting process, a high frequency of cylindrical callus formation instead of roots was obtained. To shed light on the inner anatomy of the callus chunks, we compared their cell organization and cell-wall composition to those of roots. Whereas in the root, cells were organized in circumferential symmetry, no symmetry was found in the callus. Instead, a more chaotic accumulation of meristematic-like cells with sporadic clusters of tracheary elements laid in different directions were observed. The outer cell layer of the callus often included swollen cells with thin cell walls. Most callus cells stained more strongly for cellulose and lignin than cells in the root meristem. In addition, specific antibodies to methylesterified and de-methylesterified pectin showed differential staining of callus vs. root cells indicating cell wall differences. Strikingly, roots were seen to differentiate from the chaotic cell organization of the callus, albeit at low rates. Further investigation of the cellular and molecular mechanisms underlying callus formation, are required.

Coupling of Cell Division and Differentiation in Arabidopsis thaliana Cultured Cells with Interaction of Ethylene and ABA Signaling Pathways.

Recent studies indicate direct links between molecular cell cycle and cell differentiation machineries. Ethylene and abscisic acid (ABA) are known to affect cell division and differentiation, but the mechanisms of such effects are poorly understood. As ethylene and ABA signaling routes may interact, we examined their involvement in cell division and differentiation in cell tissue cultures derived from several Arabidopsis thaliana plants: wild type (Col-0), and ethylene-insensitive mutants etr1-1, ctr1-1, and ein2-1. We designed an experimental setup to analyze the growth-related parameters and molecular mechanisms in proliferating cells upon short exposure to ABA. Here, we provide evidence for the ethylene–ABA signaling pathways’ interaction in the regulation of cell division and differentiation as follows: (1) when the ethylene signal transduction pathway is functionally active (Col-0), the cells actively proliferate, and exogenous ABA performs its function as an inhibitor of DNA synthesis and division; (2) if the ethylene signal is not perceived (etr1-1), then, in addition to cell differentiation (tracheary elements formation), cell death can occur. The addition of exogenous ABA can rescue the cells via increasing proliferation; (3) if the ethylene signal is perceived, but not transduced (ein2-1), then cell differentiation takes place—the latter is enhanced by exogenous ABA while cell proliferation is reduced; (4) when the signal transduction pathway is constitutively active, the cells begin to exit the cell cycle and proceed to endo-reduplication (ctr1-1). In this case, the addition of exogenous ABA promotes reactivation of cell division.

Brachypodium distachyon MAP20 functions in metaxylem pit development and contributes to drought recovery.

Pits are regions in the cell walls of plant tracheary elements that lack secondary walls. Each pit consists of a space within the secondary wall called a pit chamber, and a modified primary wall called the pit membrane. The pit membrane facilitates transport of solutions between vessel cells and restricts embolisms during drought. Here we analyzed the role of an angiosperm-specific TPX2-like microtubule protein MAP20 in pit formation using Brachypodium distachyon as a model system. Live cell imaging was used to analyze the interaction of MAP20 with microtubules and the impact of MAP20 on microtubule dynamics. MAP20-specific antibody was used to study expression and localization of MAP20 in different cell types during vascular bundle development. We used an artificial microRNAs (amiRNA) knockdown approach to determine the function of MAP20. MAP20 is expressed during the late stages of vascular bundle development and localizes around forming pits and under secondary cell wall thickenings in metaxylem cells. MAP20 suppresses microtubule depolymerization; however, unlike the animal TPX2 counterpart, MAP20 does not cooperate with the γ-tubulin ring complex in microtubule nucleation. Knockdown of MAP20 causes bigger pits, thinner pit membranes, perturbed vasculature development, lower reproductive potential and higher drought susceptibility. We conclude that MAP20 may contribute to drought adaptation by modulating pit size and pit membrane thickness in metaxylem.

Vessel elements of two thelypteroid ferns-part I

BackgroundHydraulically efficient xylem was evolved in the vascular plants as an apomorphy of the group. Main xylem components involved in water conduction are tracheid and vessel. Vessels, in which two ends are perforated, constituted major evolutionary innovation within vascular plants, presumably providing more efficient solute conduction. Not all vascular plants have vessels. In pteridophytes vessels are present only in seven genera. The contention lies regarding the presence and distribution of vessel in pteridophytes are the impulsive force of this investigation.MethodsTracheary elements are isolated following the standard maceration technique, then hand-razor cut longisections are passed through the aqueous alcohol grades and air-dried samples are placed on stub, sputter coated with gold and examined with SEM.ResultsTwo thelypteroid ferns viz. Ampelopteris prolifera (Retz.) Copel. and Thelypteris interrupta (Willd.) K. Iwats. are having vessel elements in root, rhizome, stipe, rachis, primary vein/costa, root-rhizome and rhizome-petiole junction i.e. through entire vascular connection of the plant body though the vessel network is interrupted and joined with parenchyma at the end in some places. Presence of vessel elements in the costa of pteridophytic taxa is first time reported by this study. Vessel end-walls are obliquely placed (root, rhizome, and stipe) but oblique to horizontal orientation is noticed in the primary vein/costa. End-walls are with simple, intermediate and compound perforation plates observed through SEM imaging as well as with tissue specific stain. Studied taxa are grown either in terrestrial microclimate of two contrasting environments i.e. sun and shade (A. prolifera) or in open swampy land (T. interrupta) with moderate to highly disturbed places as rapid proliferating populations showing interpopulation variations of tracheary elements length–width(s) and vessel end-wall length–width(s).ConclusionVessel elements are present throughout the entire vascular connections of the plant body of A. prolifera (Retz.) Copel. and T. interrupta (Willd.) K. Iwats. Interpopulation variation of tracheary elements length–width(s) and vessel end-wall length width(s) are noticed. Till date only seven genera of pteridophytes are reported for the presence of vessel and these two genera are the new addition with the previous.

Comparison of Iris L. morphogenesis in in vitro culture with seedling development in some highly specialized plants

A sharp change in the course of ontogenesis and transition to another type of nutrition (parasitic nutrition) is supposed to contribute to similarity of the morphogenetic processes in in vitro plants and seedlings of higher parasitic plants. Tracheary elements are formed in the shoots of Iris in in vitro culture as an adaptive response of the plant body to culture conditions. Underdevelopment of the vascular system for transporting nutrients from the agar medium to plant tissues results in the formation of this vascular system in the maternal shoots. Macronutrients and plant hormones are transported to the zones of meristematic activity via the vascular system. Parasitic plants obtain nutrients from their hosts through the newly formed vascular system. The elements of tracheary structures are unique for haustoria and play a vital role not only in deposition and transport of macronutrients, but also in the hydrostatic pressure regulation and protection against pathogens. The vascular system of parasitic plants is disordered similar to that of regenerated plants. This vascular system can be referred to as a distribution network, the structure capable of transforming a set of cells into the cellular transport system, which unites tissues and organs into a single growing organism.

Plant Tissue Cultures.

Plant tissue cultures are an efficient system to study cell wall biosynthesis in living cells in vivo. Tissue cultures also provide cells and culture medium from which enzymes and cell wall polymers can easily be separated for further studies. Tissue cultures with tracheary element differentiation or extracellular lignin formation have provided useful information related to several aspects of xylem and lignin formation. In this chapter, methods for nutrient medium preparation and callus culture initiation and its maintenance as well as those for protoplast isolation and viability observation are described. As a case study, we describe the establishment of a xylogenic culture of Zinnia elegans mesophyll cells.