The Control of Vascular Branching in Coleus 2. The Corner Traces

Corner trace connections are less well defined than those ofthe side bundle in Coleus, the locations of branch points, branchpartners, and number of connections made by a corner trace beingmore variable. The auxin balance between corner traces was alteredby leaf removal and by application of exogenous auxin. Branchingof new strands was shifted toward the pre-existing strand withthe lower auxin flux, but only within a narrow range of developmentalstages and with the imposition of a large auxin imbalance. Branchingoccurred only in nodal regions, as in control plants. Thus,auxin balance can be made to control xylem strand branching,but it does not account fully for the control of vascular branchingin intact plants. In the intact pattern, corner trace branchesappear to be directed toward the pre-existing strand with thehigher auxin flux. It is proposed that, in the vicinity of astrand with high flux, auxin is transported laterally withinthe nodal vascular cambium, facilitating vessel differentiationbetween strands in the derivatives of the vascular cambium.These vessels comprise the connections between traces. Coleus, vascular differentiation, vascular anatomy, vascular branching, vascular patterns, auxin, auxin balance, node     

Inhibition of xylogenesis by morphactin in pith parenchyma explants of Lactuca

Pith parenchyma explants of Romaine lettuce (Lactuca salivaLinn. var. Roman?) incubated in the dark for 7 days at 25?Con a nutrient medium containing sucrose, IAA. and kinetin exhibitedextensive differentiation of tracheary elements. The additionof CFL to the medium strongly inhibited tracheary element formation.The lack of tracheary strand formation in the CFL-treated explantssuggests the inhibition of auxin transport. Conclusive evidencethat CFL influences the anatomy of differentiating xylem elementswas lacking. The addition of CFL to various combinations ofxylogenic media was not stimulatory to xylem element formationbeyond the differentiation response observed in the absenceof CFL. Unique patterns of tracheary element formation producedby cytokinin media containing IAA, 2,4-D, and NAA, respectively,were abolished by CFL. As indicated by counts of total trachearyelements formed per explant, the addition of cysteine to a CFL-containingmedium reversed the inhibitory effect of CFL. Tracheary strandformation was not re-established in the explants cultured onthe cysteine+CFL medium. Tracheary element formation was completelysuppressed by TIBA. Cysteine had a slight effect on the inhibitionof differentiation by TIBA. These observations suggest thatCFL inhibits some sulfhydryl- containing system involved eitherin the process of xylem differentiation or in some prerequisiterole necessary for the induction of tracheary element formation. (Received December 27, 1972; )     

The Control of the Differentiation of Vascular Networks

A polar control of vascular differentiation by stimuli movingfrom the shoot to the root has been demonstrated many timesand accounts for some traits of the pattern of vascular strands.The problem dealt with here is whether this polar mechanismcontrols the formation of the vascular networks which are commonin leaves. Networks generally, though not invariably, includestrands which do not connect the shoot to the root, and thereforecannot be considered as polar. A small transverse vascular strandin Phaseolus stems includes neighbouring vessels which haveopposite shoot to root directions. A developmental study indicatedthat vessels with opposite polarities do not mature at the sametime, and suggested that there are repeated changes in the directionof the flow of the polar stimuli which control vessel formation.Experiments with Pisum show that auxin can induce the formationof a non-polar strand of xylem when the location of the sourceof auxin, and the resulting direction of auxin flow, are changedrepeatedly. It is concluded that in the first stages of vasculardifferentiation there is a determination of the axis, but notof the direction, of the movement of differentiation-inducingstimuli. When the rate of development and of the productionof stimuli is not synchronous throughout the organ, this earlydetermination of an axis of movement leads to the differentiationof vascular networks.     

The Induction of Fibre Differentiation in Peas

The problem studied in this work was that of the internal controlof the formation of strands of fibres in Pisum sativum. It isshown that fibre differentiation is dependent on stimuli originatingin young leaf primordia. Removing these primordia early enoughprevents fibre differentiation; changing the position of theleaves experimentally changes the position of the fibres aswell. It was demonstrated that some stimuli for fibre differentiationmust flow through the strands at the time they differentiate.The evidence for this flow is in experiments concerning theability of very young fibre strands to regenerate after cutsas well as in experiments concerning their pattern of joining.The stimuli which originate in the leaves and control the differentiationof fibres and xylem are shown to differ in at least one component:auxin does not cause fibre differentiation and no surgical treatments,carried out on very young tissues, caused the replacement ofpart of a strand of fibres byor xylem or vice versa.     

The Induction of Differentiation of Organized Vessels in a Storage Organ

The organized differentiation of vascular tissues was studiedin a simple system which allowed vessel members to be followedindividually. Local application of auxin to pieces of turnipstorage root resulted in differentiation of vessel and sieveelements within two days. These are normally organized in alongitudinal fashion. The induction of differentiation is inhibitedby triiodobenzoic acid. The number of differentiated cells dependedon the auxin concentration and also on the length of time thetissue was allowed no differentiate. No vessel members wereobserved in less than 48 h and the minimum effective IAA concentrationwas 8 x 10^–6 M. The results established a simple, quantitativesystem for the study of vessel differentiation. Brassica campestris cv. Rapifera, auxin, differentiation, storage root, vessel, xylem     

Foliar and Axial Aspects of Vascular Differentiation: Hypotheses and Evidence

A comparison is made between foliar and axial vascular differentiation. Current thoughts and new evidence are presented on the role of hormones in controlling the differentiation of vascular tissues in organized and tumorous tissues, focusing on the role of auxin and cytokinin in controlling phloem and xylem relationships, vessel size and density, cambium sensitivity, vascular adaptation and xylem evolution in deciduous hardwood trees. The possible role of wounding is also considered. A new hypothesis, namely, the leaf-venation hypothesis, is proposed to explain the hormonal control of vascular differentiation in leaves of dicotyledonous plants. Experimental evidence in support of the hypothesis is presented showing that hydathodes, the water-secreting glands, are the primary sites of auxin synthesis during leaf morphogenesis. Vessel element patterns similar to those found in hydathodes were experimentally induced by exogenous auxin application.     

A Microautoradiographic Study of Auxin Transport in the Stem of Intact Pea Seedlings (Pisum sativum L.)

Soluble-compound microautoradiography was used to determinethe distribution of radioactivity in transverse sections ofintact dwarf pea stems (Pisum sativum L.) following the applicationof [³H]IAA to the apical bud. Near the transport front labelwas confined to the cambial zone of the axial bundles, includingthe differentiating secondary vascular elements. Fully differentiatedphloem and xylem elements remained unlabelled and no radioactivitywas detected in the leaf or stipule traces. Similar resultswere obtained in experiments with Vicia faba L. plants. Nearerthe labelled apical bud of the pea there was a more generaldistribution of label and evidence was found of free-space transportof radioactive material in the pith. When [³H]IAA was applied to mature foliage leaves the greatestconcentration of label was found in the differentiated phloemelements of the appropriate leaf trace and in the phloem ofthe adjacent axial bundles. Both basipetal and acropetal transportwas detected in this case. These results are consistent with the conclusions drawn fromearlier transport experiments which indicated that in the intactplant the long-distance basipetal transport of auxin from theapical bud takes place in a system which is separated from thephloem transport system and suggests that the vascular cambiumand its immediate derivatives may function as the normal pathwayfor the longdistance movement of auxin in the plant. The physiologicalsignificance of such a transport system for auxin is discussed.     

The Vascular Pattern of Italian Ryegrass (Lolium multiflorum Lam.): 3. The Leaf Trace System, and Tiller Insertion, in the Adult

The leaf trace system in the region of congested internodesat the base of Lolium multiflorum is described. A typical major trace in a leaf consists of a collateral bundlehaving a double bundle sheath and incorporating a certain amountof sclerenchyma. As such a leaf trace is followed down intothe stem it increases in diameter, loses the inner (mestome)bundle sheath, and the xylem becomes associated with xylem transfercells. Lower down, the bundle diameter is reduced although nowit has become amphivasal. The internal xylem only is still associatedwith transfer cells. The proximal portions of the bundle aremuch reduced, transfer cells, mestome sheath, and sclerenchymaare lacking and the now insignificant bundle merges with a lowerleaf trace or some other vascular tissue. Such a bundle in thestem may be in direct contact via bridges with other leaf traces,with the nodal plexus, and with the peripheral plexus that surroundsthe inner leaf trace system. In the base of a typical young plant, approximately one-halfof all leaf traces, including all the median veins, join bundlesfrom the next oldest leaf. Approximately one-third join thenodal plexus, and the remainder variously join bundles fromthe same or next but one oldest leaf to join the peripheralplexus. The differentiation of tiller insertions into the pre-existingmain stem system is highly variable. In a very young tillera number of traces were seen to terminate before the main systemwas reached suggesting basipetal differentiation. The actualconnections made by the tiller traces may occur with any nearbyleaf trace, the nodal plexus, or with the peripheral plexus.Later differentiating leaf traces in a tiller join leaf tracesof the tiller itself. Occasional bundles from secondary tillers by-pass the vasculartissue of the primary tiller to join directly with that of theparent plant. Vascular connections between parent and tiller,although very variable, appear to be totally comprehensive froma functional standpoint.     

Organogenesis, Differentiation and Histolocalization of Alkaloids in Cultured Tissues and Organs of Duboisia myoporoides R. Br.

Duboisia myoporoides R. Br. shoots were regenerated from non-organogenicand organogenic calli induced with nine different cytokinin/auxincombinations. Alkaloid colour reagents localized tropane alkaloidsin the vascular regions which had large cells in the secondaryxylem of the basal stem sections of the non-rooted shoots. Tropanealkaloids were localized in shoots regenerated from calli inducedwith two different cytokinin/auxin combinations. No alkaloidswere localized in shoots regenerated from calli induced withother cytokinin/auxin combinations. However, only nicotine wasdetected by GC-MS in the non-rooted shoots regenerated fromcalli induced with three different cytokinin/auxin combinations.Tropane alkaloids were also localized in xylem cells of rootsregenerated from calli induced with two different cytokininand auxin combinations independently. The presence or absenceof nicotine, hyoscyamine and scopolamine in different culturedplant materials was confirmed by GC-MS, indicating that althoughthe root is the main site for alkaloid biosynthesis, with suitablecell differentiation, alkaloid biosynthesis may take place incultured shoots without root initiation. Copyright 2000 Annalsof Botany Company Duboisia myoporoides, Corkwood tree, Solanaceae, tropane alkaloid, alkaloid localization, shoot culture, root culture, iodoplatinate     

Auxin in the Cambium and its Differentiating Derivatives

Cambium and differentiating xylem and phloem tissues from thetrunks of trees of Acer pseudoplatanus L., Fraxinus excelsiorL., and Populus tremula L. were extracted with ether and testedfor auxin, which was found on chromatograms of the acidic fractionat an Rf corresponding to that of indol-3yl-acetic acid in fivesolvent systems. In addition, small amounts of auxin with ahigher Rf in ammoniacal isopropanol were found in phloem samples.The amounts of auxin were greatest in xylem samples, less inthe cambium, and least in phloem. The differences, which cannotbe explained in terms of differential losses during extractionand purification, suggest that auxin is actually formed in differentiatingxylem tissue. The significance of these results is discussed.     

The Initiation and Development of Secondary Xylem in Axillary Branches of Populus deltoides

The initiation of secondary xylem in elongating axillary branchesof Populus deltoides Bartr. ex Marsh. is independent of thatin the main stem. Although secondary xylem differentiates acropetallyin the main stem, it does not differentiate from the stem intothe axillary branch. Secondary xylem is usually initiated ininternode 4 (occasionally 3) of the axillary branch, and fromthis site it develops both acropetally in the elongating branchand basipetally toward the main stem. Secondary vessel differentiationalways precedes fibre differentiation. Although secondary xylemdifferentiates in internodes that have ceased elongation, itdifferentiates first in traces of the vascular cylinder servingrapidly expanding and maturing foliage leaves. As younger leaveson the branch expand and mature, secondary xylem differentiatesin their traces eventually producing a complete secondary vascularcylinder. Scale leaves do not initiate secondary xylem independentlyin their traces; they are activated by adjacent traces in thevascular cylinder serving foliage leaves. Once established,the primary-secondary vascular transition zone advances acropetallyin a branch just as it does in the main stem. Populus deltoides Bartr. ex Marsh., cottonwood, axillary branches, secondary xylem, plastochron index, post-dormancy development, xylem.     

Regeneration Around Wounds and the Control of Vascular Differentiation

The question which was the basis of this work was whether (a)vascular regeneration around wounds includes a replacement ofdamaged tissues or (b) only new vascular strands, which arenormally formed from the cambium, are diverted around wounds.It was found that in Coleus and Cucumis no connections are formedto damaged sieve tubes and vessels, so that their continuityaround wounds is not restored. Pisum plants were wounded underconditions in which growth could not be influenced and the areaof the xylem in cross-section was measured 1 month later. Thewounds, which damaged the vascular tissues, significantly increasedvascular differentiation, indicating the replacement of a longnon-functional region of damaged tissues. The results indicatethat in the intact plant vascular differentiation is controllednot only by stimuli from the leaves but also by the capacityof the mature vascular system to transport these stimuli.     

Polarity and the Induction of Organized Vascular Tissues

This work deals with those properties of plant tissues whichare responsible for the organization of vascular cells in orderedstrands. It is shown that auxin alone is sufficient to causethe differentiation of strands of xylem cells in the parenchymaof pea roots. An artificially induced strand, once it is formed,attracts towards itself newly induced vascular strands, andthis attraction results in the union of old and new strands.It is also shown that the application of auxin to natural vasculartissues prevents their being joined by newly induced vascularstrands. It is proved that this is dependent on a directionaleffect and not simply on a local accumulation of auxin. To understand these results, it must be assumed that the polarityin terms of auxin transport is increased during the processof vascular tissue induction. The same polarity, once established,is maintained by the presence of auxin, so that the differentiationof strands perpendicular to the axis of this polarity is prevented.These characteristics of plant tissues concerning auxin transportexplain the basic phenomena of the organization of vascularcells in defined and ordered strands.     

ANATOMY OF NODES VS. INTERNODES IN COLEUS: THE NODAL CAMBIUM

Secondary growth begins in the nodal regions before the internodal regions in Coleus, so that longitudinally discontinuous vascular cambia are formed in the 6th through the 9th or 10th nodes, where the internodal cambium becomes continuous between nodal cambia. The nodal cambia are identifiable by radial seriation in interfascicular regions, typical cytology of fusiform initials, and the presence of a ray system. Anatomical features distinct from the primary plant body are shared by the nodal and internodal cambia. Branching of primary vascular strands, restricted to procambium and phloem, is virtually confined to nodal regions. In secondary growth, vascular branching of xylem and phloem occurs in both nodes and internodes. Xylem strand branches are formed only from derivatives of vascular cambia. It is proposed that the cambium provides the secondary plant body an efficient channel for lateral auxin transport, by which branching across interfascicular regions is facilitated.     

Phloem-associated auxin response maxima determine radial positioning of lateral roots in maize

In Arabidopsis thaliana, lateral-root-forming competence of pericycle cells is associated with their position at the xylem poles and depends on the establishment of protoxylem-localized auxin response maxima. In maize, our histological analyses revealed an interruption of the pericycle at the xylem poles, and confirmed the earlier reported proto-phloem-specific lateral root initiation. Phloem-pole pericycle cells were larger and had thinner cell walls compared with the other pericycle cells, highlighting the heterogeneous character of the maize root pericycle. A maize DR5::RFP marker line demonstrated the presence of auxin response maxima in differentiating xylem cells at the root tip and in cells surrounding the proto-phloem vessels. Chemical inhibition of auxin transport indicated that the establishment of the phloem-localized auxin response maxima is crucial for lateral root formation in maize, because in their absence, random divisions of pericycle and endodermis cells occurred, not resulting in organogenesis. These data hint at an evolutionarily conserved mechanism, in which the establishment of vascular auxin response maxima is required to trigger cells in the flanking outer tissue layer for lateral root initiation. It further indicates that lateral root initiation is not dependent on cellular specification or differentiation of the type of vascular tissue.     

The Vascular Pattern of Italian Ryegrass (Lolium multiflorum Lam.) 1. The Young Seedling and Its Variability

The vascular system of the young Italian ryegrass seedling (Loliummultiflorwn Lam.) is very similar to that of oat (Avena) whichhas been described in the past. Early accounts however failto record the distinctive separate course of the xylem and phloemconstituents of the bundles at the coleoptilar node. The ascendingmesocotylar trace is bicollateral. It continues up, in part,into the mid-vein of the first foliage leaf and arches over,joining the bases of the two coleoptilar traces, to become thecollateral scutellar trace. Considerable variation exists from seedling to seedling in thedetails of the vascular pattern. This is coupled with additionalvariation in mesocotyl elongation, extent of the coleoptilarnode, number and location of lateral root primordia and theirmaturation, rate of growth of coleoptile and first foliage leavesand the rate of vascular differentiation within them. Possiblecauses of this variability are discussed and it is noted thatapparently identical plants, selected on a single morphologicalcriterion such as degree of elongation of the coleoptile, maybe expected to differ markedly in other morphological and anatomicalaspects, influencing their performance in physiological experimentation.     

Vascular and General Anatomy of the Rootstocks of Three Stemless Viola Species

The primary vascular system of the rootstock of Viola rotundifolia,V. odorata and V. cucullata consists of an open system of threesympodia, corresponding to three orthostichies (parastichies)of leaves in the 1 /3 phyllotaxy. Between the major, collateralbundles there are vascular strands consisting of only primaryphloem. The vascular supply to axillary buds developing in thefirst season of growth is the same in flowering and vegetativebuds and there is a homology between the bracts on the pedunclesof axillary flowers and the prophylls of the axillary vegetativebranches. The overwintering portions of the rootstocks are somewhatwoody. The uneven development of secondary xylem correspondsto the locations of the sympodia. Secondary xylem closes a leafgap of a median trace only slowly, but any gap in the phloicsystem is closed quickly by secondary growth. Viola rotundifolia, V. odorata, V. cucullata, rootstock, vascular anatomy     

Naturally Occurring Regenerative Differentiation of Xylem Around Adventitious Roots in Luffa cylindrica Seedlings

Regeneration of xylem induced by adventitious root formationin the hypocotyl of Luffa cylindrica Roem. seedlings is described.This naturally occurring form of xylem regeneration involvesthe formation of a bypass of regenerated tracheary elementsaround a root without external severance of the vascular strands.The regeneration of xylem around an adventitious root is polarand is very similar in its developmental pattern to the well-knownxylem regeneration induced by wounding vascular strands. Adventitious root formation, Luffa cylindrica Roem, regenerated tracheary elements, vascular differentiation, xylem regeneration