Development and Structure of Unusual Xylem Elements During Graft Union Formation in Picea sitchensis L.

Electron microscope studies of Sitka spruce have been undertakento observe the sequence of cellular development following grafting.Observations at the graft interface reveal a recovery and regenerationsequence associated with union formation. Xylem elements differentiateddirectly from the vascular cambia of the rootstock and scionare different from elements arising from parenchymatous callusderived from ray parenchyma, which may be produced in an attemptto establish a connecting water conducting system as rapidlyas possible. Failure to form such a system may be a primarycause of graft failure. Grafting, Picea sitchensis, xylen elements     

The Effect of Scion Water Potential on Graft Success in Sitka Spruce (Picea sitchensis)

The effect of scion storage on graft success in Sitka sprucehas been examined. Scions kept in cold store for up to 7 d werefound to graft successfully to active rootstocks in about 95%of cases. From 7 d, however, there was a marked decline in successuntil, after 14 d of storage prior to grafting less than 40%of grafts survived. Scion leaf-water potential decreased continuouslyduring storage and fell dramatically during the first 3 d aftergrafting. In successful grafts, scion water potential then recovered,reaching a stable level after 3 weeks. No recovery occurredin grafts, which eventually failed. The relationship betweenscion leaf water potential and graft success rate is discussedwith reference to the role of callus development in graft formation. Picea sitchensis, water potential, graft success     

The Anatomy of the Developing Bud Union and its Relationship to Dwarfing in Apple

Light microscopy has been used to study the effect of dwarfingand semi-dwarfing apple rootstocks on the early developmentof bud-unions with 'Gala', and the anatomy of 2-year-old bud-unionsbetween 'Bramley' and the same rootstocks. The bridging of thecut edges of the cambia of bud and rootstock was achieved bydifferentiation of callus formed at an early stage in budding.New cambial cells were aligned at right angles to the pre-existingcambia, with their long axes horizontal. Subsequently-formedxylem adopted this arrangement, so that fibres and vessels werearranged obliquely to the axis of the stem. At the interfacebetween the bud and dwarfing rootstocks vessels with smallerthan normal diameter were formed, indicating the presence ofelevated levels of auxin in this region. In addition, littlexylem was produced in the adjacent rootstock tissue. In thecase of semi-dwarfing rootstocks, the rootstock produced normalxylem after a brief interruption. We suggest that failure ofauxin to cross the bud-union interface in the case of the dwarfingrootstocks leads to reduced rootstock xylem formation, and hencea poor supply of water and minerals to the scion, and this underliesthe dwarfing effect.Copyright 1994, 1999 Academic Press Apple, budding, dwarfing, anatomy, graft union     

The effect of applied heat on graft union formation in dormant Picea sitchensis (Bong.) Carr

The effect of application of heat on the development of graftunions in dormant Picea sitchensis (Bong.) Carr (Sitka spruce)was observed at monthly intervals from October to March. Noadvantage over controls was gained in terms of final graft successrates. It is suggested that this may have been a result of themild winter conditions during the course of the experiment,which enabled callus formation to occur in unheated controls.On the other hand, scions of grafts which had been heated for3 weeks, and then returned to ambient conditions showed considerablygreater growth than unheated controls during the following springand summer. The amount of extension depended on how late inthe winter the graft had been prepared and heated, with leastgrowth by grafts made in October, and most by grafts made inMarch. Microscopical examination showed that callus formationwas more rapid in heated grafts than in controls, although callusformation occurred in all control grafts examined. Heat alsoinduced cambial reactivation and tracheid formation, particularlyin the vicinity of needle or bud traces in the scion. In keepingwith this, it was found that the presence of needles and budswas essential for cambial reactivation in normal dormant stemsand that reactivation was restricted to the heated region. Key words: Grafting, Sitka spruce, callus, dormancy, differentiation     

The Structure and Composition of Bead-like Projections on Sitka Spruce Callus Cells Formed during Grafting and in Culture

Projections at the cell wall surface in callus formed at thegraft interface and in vitro in Picea sitchensis (Bong.) Carrhave been examined using light and electron microscopy. Thesestructures range in size from 1 to 5 µm in diameter, andconsist of a homogeneous matrix enclosing one or more fibrillar/vesicularcores. Histochemical and cytochemical studies have shown thatthe homogeneous matrix is made up of a mixture of pectins, carbohydrate,protein and fatty acids, while the fibrillar/vesicular componentis mainly carbohydrate and pectins. The possible functions ofthese structures are discussed on the bases of their composition.Copyright1993, 1999 Academic Press Picea sitchensis, callus, cell-wall projections, histochemistry, cytochemistry     

<Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> as a Model System for Graft Union Development in Homografts and Heterografts

Homografting of Arabidopsis thaliana scions on stocks of A. thaliana and heterografting on other species were used to study the compatibility and the ontogeny of graft union formation. Highly compatible homografting with scions of young leafy inflorescence stems was obtained on stocks of inflorescence stems growing from large 3-month-old A. thaliana plants. Histologic analysis revealed four developmental stages of graft union formation in Arabidopsis homografting: (1) development of a necrotic layer, (2) callus proliferation in the grafted scion, (3) differentiation of new vascular tissues within the scion, and (4) a full vascular graft union formation between the scion and the stock. Vascular connections were formed within the callus bridge between rootstocks and scions 15 days after grafting. Heterografts of Arabidopsis on two members of Brassicaceae, cabbage (Brassica) and radish (Raphanus), showed partial incompatible interaction with a lower level of vascular differentiation. Arabidopsis grafting on tomato (Solanaceae) rootstock showed complete incompatibility and limited noncontinuous differentiation of new vascular tissues that did not cross the scion/stock boundary. Although lacking scion/stock vascular connections, Arabidopsis scions grafted onto tomato rootstock flowered and produced seeds. This may indicate some nonvascular functional connections between the two plants, probably of parenchyma cells, further emphasizing the usefulness of Arabidopsis as a model plant for studying various levels of the complicated scion/stock relationships expressed in grafting biology. Experiments with dye transport in the xylem showed that although in general there was an agreement between the histologic study and dye transport, in Arabidopsis homografts water transport frequency was lower than functional and histologic compatability. We conclude that homografting and heterografting of Arabidopsis inflorescence stems is a convenient and reproducible method for studying the fundamental cellular genetic and molecular aspects of grafting biology.     

The Anatomical Relationship Between Cambial Regeneration and Root Initiation in Wounded Winter Cuttings of the Apple Rootstock M.26

Root primordia differentiate remote from the existing vasculartissue of split- or incision- wounded winter cuttings. Thisis preceded by heavy callusing of the wounds and basal cut surface.Most of the callus is cortical in origin but callus is alsoformed as a result of damage to the cambium. In the incisiontreatment a new cambium differentiates within the cambial callusfrom the undamaged cambium on either side so as to form an outward-pointingsalient enclosing randomly orientated xylem. Two such salientsare formed in the split base treatment since each resultanthalf behaves like a separate cutting. The salients are subdividedhorizontally into finger-like projections due to the dispositionof the rays at the edges of the wound. Once a root primordiumforms, differentiation of procambium-like tissue between itand one or more projections proceeds rapidly, followed by outgrowthof the root. Roots emerge from the basal callus and in verticalfiles from the wound callus. The split base treatment increases16-fold the number of rooted cuttings over controls, while incisionwounds increase rooting four-fold; this is associated with thefact that more rhizogenic (salient-forming) sites are formedby splitting the base. The physical and anatomical factors involvedin cambial regeneration are discussed. Anatomy, apple, callus, cambium, cambial regeneration, M.26 rootstock, Malus pumila L., rooting, root initiation, wound response     

GRAFT UNION FORMATION IN DOUGLAS-FIR

Greenhouse-grown Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) graft unions were examined between 2 and 84 days after grafting. Room temperature was maintained at 60-70 F throughout the growing season. In most respects grafts of Douglas-fir followed development patterns previously reported for spruce and pine grafts, but specific differences were noted in contributing cell types, time of formation, and mode of healing. The time interval from first occurrence to occurrence in 80% of the grafts is as follows: contact layers, 2 days; callus bridges, 10-14 days; periderm, 10-17 days; cambia, 17-23 days. Callus bridges were generally of secondary phloem or cortex origin. Callus lignification began along cut edges of the union at 14 days and was completed across the entire length of the union by 17 days. Lignified tracheids were continuous across union zones at 35 days. When proper grafting techniques were used, all tissue systems necessary for a successful union were present 35 days after grafting. Poor grafting techniques at times retarded cambium formation for 3 months or more.     

Secondary Growth in Bougainvillea

The anomalous secondary growth was studied in roots and stemsof two species of Bougainvillea. The anomalous cambia arisesuccessively in centrifugal order, each originating among thederivatives of the preceding cambium. Each cambium layer functionsbidirectionally producing xylem towards the inside of the axisand phloem towards the outside. The sequence of production ofvascular cells is the following: (1) conjunctive tissue andxylem fibres towards the inside; (2) phloem towards the outside;(3) additional xylem with vessels towards the inside and additionalphloem towards the outside. The new cambia arise outside theoldest phloem cells of a given increment. This phloem may benonfunctional and crushed at that time. The phloem and the xylemdifferentiate from radially seriated derivatives produced sequentiallyby tangential divisions in the cambium. Divisions among thephloem initials and growth readjustments in the differentiatingxylem obscure the radial seriation to a moderate extent.     

Prevention of auxin-induced vascular differentiation in wound callus by surface-to-surface adhesion between calluses of stock and scion in cactus grafts

The relationship between formation of CVB, which connects vascularcut ends of stock and scion, and the condition of tissue whereCVB is expected to be induced was studied using a cactus graftcombination of Notocactus submammulosus var. pampeanus youngseedlings on Hylocereus trigonus plants. When a scion with its basal part removed by a transverse cutwas put onto a transverse cut surface of a stock with a distanceof 1 mm between the vascular cut ends of both graft components,CVB was formed in only 13% of the grafts in the callus massmade by adhesion between calluses of stock and scion. The percentagearose to ca. 100% when 100 ppm NAA was applied to the scionapices. This promotive effect of NAA decreased with increasingtime from grafting to NAA application. When wound calluses of stock and scion were prevented from contactonly at the portion between the vascular cut ends of stock andscion by introducing a piece of aluminium foil into the graftunion, CVB was formed in 80% of the grafts in the callus ofthe stock without NAA application. In the case where the scionwas grafted at right angle onto an additional longitudinal cutsurface made 1 mm from the vascular bundle of the stock withremaining intact parenchyma between this cut surface and thevascular bundle of the stock, CVB was not formed even when NAAwas applied. These results suggested that wound response wasa prerequisite for CVB differentiation and that surface-to-surfaceadhesion of calluses inhibited CVB formation by making calluscells differentiate into quiescent parenchyma cells rapidly.The mechanism of this phenomenon is discussed. (Received September 29, 1977; )     

A Study of the graft union in in vitro micrografted apple

Light microscopy was used to study graft union formation in in vitro micrografts of tissue cultured apple (Malus domestica. Borkh). Micrografts were constructed using horizontal incisions to form the grafting surface, and placing the cut ends of rootstock and scion into sterile silicon tubing to permit graft formation to occur.The outer morphological and histological development was similar for different stock-scion micrograft combinations but graft union formation was slower in heterografts than in autografts. Initial leaf expansion at the scion shoot apex occurred in all micrografted plantlets within 1–4 days and was not indicative of graft success. Progressive scion growth and development could be used as an indication of graft success by ten to fourteen days after grafting and probably was related to establisment of cell to cell contact at the graft interface. Microscopy showed initiation of callus proliferation in the vascular cumbium and the pith ten days after grafting. Differentiation was observed subsequently and this was reflected in scion development. Longitudinally orientated cambial cells began to differentiate between twenty and forty days after grafting, and formed a bridge between the vascular cylinders of scion and rootstock. The scions at this stage had as many as eight newly expanded leaves and micrografts were strong enough to permit silicon sleeve removal without damage. Continuity of new vascular elements in rootstock and scion was established around forty days. New vascular elements curved slightly in towards the pith to form a c shaped bridge across the graft union. Vascular development continued until it reached completion after six months.     

The Developmental Anatomy of the Root System in Yam Bean, Pachyrhizus erosus Urban

Investigations revealed that the anatomy of the primary radicularroot of yam bean (Pachyrhizus erosus L.) was typically dicotyledonousexcept that the xylem was not completely developed centripetally.Most of the roots had tetrarch xylem, although a few triarchand pentarch roots were also observed. In both tuberous andnon-tuberous roots, secondary thickening occurred by the formationof the meristematic vascular cambium which formed secondarytissues in a normal fashion. Subsequently, tuberization wasinitiated in the secondary xylem by the development of anomalous‘secondary’ cambia from parenchyma cells surroundingvessel elements. Anomalous ‘secondary’ cambia alsodeveloped from parenchyma cells not associated with vessels.Subsequently, anomalous ‘tertiary’ cambia differentiatedfrom tissues produced by the anomalous ‘secondary’cambia. Activities of these anomalous cambia resulted in theproduction of parenchyma storage cells and were chiefly responsiblefor the growth of the mature tuber. Pachyrhizus erosus L., yam bean, tuberous root, anatomy, anomalous ‘secondary’ cambia, anomalous ‘tertiary’ cambia, centripetal xylem development     

Graft establishment between compatible and incompatible Prunus spp

Graft compatibility has been studied in apricot (prunus armeniacaL.) grafted on Prunus cerasifera Ehrh. Two apricot cultivars,one compatible and one incompatible on this rootstock, wereselected for this study. In these species incompatibility isonly manifested by tree breakdown at a late phase of the tree'slife. The process of graft union formation was observed forthe first month following grafting. No differences were foundeither in the process of healing or in its kinetics. Thus, callusproliferation, callus differentiation and vascular connectionsare established in the same way and at the same time in bothcompatible and incompatible grafts. However, clear differencesexist in the level of differentiation of the callus produced.While in compatible grafts, callus quickly differentiates intocambium and vascular tissue, in incompatible grafts this differentiationis not complete and a portion of the tissue evolves into a parenchymatoustissue that coexists with the differentiated vascular tissue. Key words: Graft, Prunus, compatibility     

The Effect of a Grafted Bud on Vascularization of the Petiole in Phaseolus, Datura and Lycopersicon

The radially-organized petiole of Phaseolus and the dorsiventralpetioles of Datura and Lycopersicon were used as stocks in budgrafting. Petiolar structure was examined after some weeks'growth of the scion, during which the grafted petiole came tofunction as a stem in supporting normal, vigorous shoot growthand was not abscised even at the end of the growing season.Below the graft union, reactivated petiolar cambium producedmassive amounts of secondary tissue with greatly enlarged vessels.Cambial activity was confined to the existing vascular bundlesexcept for the development of a little inter-fascicular cambiumin young, grafted Phaseolus petioles. Datura petioles woundedbelow the graft union, by a cut into the petiolar are from eitherthe abaxial or the adaxial surface and removal of a 1 cm lengthof tissue, responded by restoring the vascular are (abaxialwounds) or almost completing a vascular ring (adaxial wounds).In grafted, wounded Lycopersicon petioles the presence of deadxylem caused the separation of cambium regenerated from thecut vascular are and that arising in relation to the centralwound surface. A similar response, in which stimulated internalphloem plays an important part, occurred in certain woundedLycopersicon stems. The results are discussed in terms of thegradient induction hypothesis. graft, petiole, wound, cambium, xylem, phloem, Phaseolus multiflorus, Datura stramonium, Lycopersicon esculentum, bean, thornapple, tomato     

The position of regenerating cambia: auxin/sucrose ratio and the gradient induction hypothesis.

To account for the positions in which vascular cambia regenerate in wound callus, a gradient induction hypothesis was proposed in 1961 in terms of gradients in 'some factor as yet unknown'. It now seems likely that the gradient is based on morphogen diffusion between source and sink on opposite sides of existing cambia, with morphogen diffusing into the adjoining wound callus. It is specifically proposed that there are two morphogens, auxin diffusing centrifugally and sucrose diffusing centripetally. The cambium then regenerates along a path where the ratio of auxin to sucrose concentration is similar to that at the original cambium, and its orientation (as regards xylem and phloem formation) is determined by the direction of the gradient in this ratio. These proposals are supported by published evidence on auxin and sucrose concentration gradients across the cambium, and on their sources, movements, and known effects on vascular differentiation. Simulations of the proposed positional control system predict patterns of cambial regeneration and orientation corresponding to those observed in four different types of wound and graft.     

Callus Formation at Graft Interface in Ginkgo biloba L.

The early stages of graft union, when male branch was grafted onto female branch in Ginkgo biloba L. by cleft graft, have been observed under light microscope in order to determine the origin of callus cells between the stock and scion. Pith parenchyma cells near the graft interface were the earliest cells in response to such method of grafting. These cells dedifferentiated and then divided within 7～ 12 days after grafting. A large number of callus cells extended from the pith into the space between the graft interface linking the stock with the scion about 18～ 20 days after grafting; and then continued to proliferate and extend outwards along the space. Cambium cells and immature vascular tissue near the graft interface dedifferentiated into callus rather late. Theover all link between the stock and scion was completed in the sites 30 days after grafting. Callus cells were also produced from corticai parenchyma cells, but they were much limited in quantity. In conclusion, the graft interface may be considered as a "natural culture bed" after grafting, in which all undamaged, living cells are capable of dedifferentiation and producing callus cells for compatible graft union. In the case of G. biloba (male/female) it were the pith parenchyma cells that appeared first to form the callus cells and later extend to link the stock with the scion.     

An Early Cytochemical Marker of Commitment to Stelar Differentiation in Meristems from Dicotyledonous Plants

A cytochemical study of root apices from Vicia faba and Pisumsativum showed esterase activity to be present in the stele,root cap and rhizodermis, but almost completely absent fromthe developing cortex and quiescent centres. The meristem cellsgiving rise to the cortex were almost negative whilst thosegiving rise to the stele were positive for esterase activity.Cambia from roots, shoots and petioles of a number of dicotyledonousspecies were all positive for esterase activity. It is proposedthat esterase activity may be used as an early marker of commitmentto differentiation into stele in roots of dicotyledonous plants,and that the cambia are fully committed meristems. Pisum sativum L., Vicia fabaL., garden pea, broad bean, meristems, stelar differentiation, esterase activity, xylem differentiation, cytochemistry, cambium     

Ultrastructural Aspects of Graft Incompatibility Between Pear and Quince In vitro

Callus cells of pear (Pyrus communis cv. ‘Bartlett’)underwent lethal cellular senescence in response to graftingwith callus cells of quince (Cydonia oblonga cv. ‘VanDeman’). Similar responses occurred when the cells werein direct contact and when they were separated by a porous membranefilter. These results indicate that direct cellular contactis not necessary to elicit graft incompatibility between pearand quince. Cydonia oblonga, grafting, incompatibility, pear, Pyrus communis, quince     

Radial secondary growth and formation of successive cambia and their products in Ipomoea hederifolia L. (Convolvulaceae)

Ipomoea hederifolia stems increase in thickness using a combination of different types of cambial variant, such as the discontinuous concentric rings of cambia, the development of included phloem, the reverse orientation of discontinuous cambial segments, the internal phloem, the formation of secondary xylem and phloem from the internal cambium, and differentiation of cork in the pith. After primary growth, the first ring of cambium arises between the external primary phloem and primary xylem, producing secondary phloem centrifugally and secondary xylem centripetally. The stem becomes lobed, flat, undulating, or irregular in shape as a result of the formation of both discontinuous and continuous concentric rings of cambia. As the formation of secondary xylem is greater in one region than in another, this results in the formation of a grooved stem. Successive cambia formed after the first ring are of two distinct functional types: (1) functionally normal successive cambia that divide to form secondary xylem centripetally and secondary phloem centrifugally, like other dicotyledons that show successive rings, and (2) abnormal cambia with reverse orientation. The former type of successive rings originates from the parenchyma cells located outside the phloem produced by previous cambium. The latter type of cambium develops from the conjunctive tissue located at the base of the secondary xylem formed by functionally normal cambia. This cambium is functionally inverted, producing secondary xylem centrifugally and secondary phloem centripetally. In later secondary growth, xylem parenchyma situated deep inside the secondary xylem undergoes de‐differentiation, and re‐differentiates into included phloem islands in secondary xylem. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 30–40.     

油茶芽苗砧嫁接口愈合过程解剖学研究

采用石蜡切片法对油茶（Camellia oleifera）芽苗砧嫁接口的愈合过程进行组织解剖学研究。观察结果表明：（1）在嫁接后的第4天,嫁接口产生隔离层;在嫁接后的第8天,嫁接口的砧木产生愈伤组织;在嫁接后的第16天,嫁接口的接穗产生愈伤组织;在嫁接后的第22天,嫁接口的砧木与接穗连接;在嫁接后的第29天,嫁接口的形成层分化形成;在嫁接后的第35天,嫁接口的愈伤组织维管束形成,接穗连接成功。（2）油茶芽苗砧嫁接部位愈伤组织形成前,芽苗砧木的解剖结构在短时间内加速发育,逐渐与接穗的组织结构相似。（3）嫁接口的形成层和其它薄壁细胞组织均能产生愈伤组织,但形成层是其主要来源。