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Review
Abstract
The mechanism of graft incompatibility is not yet fully understood and many reports focus on
this problem in order to understand the mechanisms of graft development. These reports refer to
both cytological and biochemical responses occurring at an early phase in response to grafting, as
well as the consequences of these events on the future graft response. Some theories argued the
possibility that the phenomena of cellular recognition could be involved in the development of a
functional vascular connection since the first callus are formed. However, callus formation can be a
passive response to a wound with no implications in the future compatibility responses. We sum up
different reasons that may have an influence on graft success: inherent system of cellular
incompatibility, formation of plasmodesmata, vascular tissue connections, and the presence of
growth regulators and peroxidases. In addition, phloem-mobile proteins have been reported that
cross the graft interface when graft bridging is established and it is functional. This review provides
an overview of the graft response and recent advances in the knowledge of the mechanism involved
in the early responses to grafting for the development of cohesion between the stock and scion
during graft ontogeny.
# 2005 Elsevier B.V. All rights reserved.
Keywords: Cellular responses; Graft-compatibility; Plasmodesmata; Proteins; Wounding
* Corresponding author. Tel.: +34 976 71 63 12; fax: +34 976 71 63 35.
E-mail address: perrea@aragob.es (P. Errea).
0304-4238/$ see front matter # 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.scienta.2005.04.003
Contents
1.
2.
3.
4.
5.
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Callus formation: the beginning of the graft process . .
Plasmodesmata and its role in cellular communication .
Vascular connections in the graft process . . . . . . . . . .
Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
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1. Introduction
Plant grafting is a widely used means of plant propagation and growth control that is of
considerable importance in the adaptation of interesting cultivars in appropriate areas. The
grafted partners can belong to the same species or genus, but usually components that are
more genetically divergent are used. In these cases the stock and scion do not always
constitute a successful graft and show their disagreement in the form of incompatibility.
Several authors have defined the sequence of structural events during the healing of the
grafts in woody and herbaceous plants. Hartmann et al. (2002) overview in their review of
the sequence of events.
1. Cut scion tissue capable of meristematic activity is brought into secure, intimate contact
with similarly cut stock tissue in such a manner that the cambial regions of both are in
close proximity in order to interconnect through the callus bridge. Once the two
components of the graft, stock and scion, are in intimate contact, new parenchymatous
cells proliferate from both stock and scion producing the callus tissue that soon
intermingles and interlocks, filling up the spaces between the two components
connecting the scion and the rootstock.
2. New cambial cells differentiate from the newly formed callus, forming a continuous
cambial connection between rootstock and scion. Furthermore, prior to the binding of
vascular cambium across the callus bridge, initial xylem and phloem may be
differentiated. The wound-repair xylem is generally the first differentiated tissue to
bridge the graft union, followed by wound-repair phloem.
3. In the last step of the graft establishment process, the newly formed cambial layer in the
callus bridge begins typical cambial activity forming new vascular tissues. Production
of new xylem and phloem thus permits the vascular connection between the scion and
rootstock. For the majority of authors, this is considered as the basic requirement for a
successful graft (Moore and Walker, 1981a,b; Yeoman, 1984; Tiedemann, 1989).
The mechanism, in which incompatibility is expressed, is not clear and several
hypotheses have been advanced in an attempt to explain incompatibility. The majority of
hypothesis referred to an early stage of development has been related to herbaceous
systems. However, few studies have been made on early establishment in woody plants,
where in many cases incompatibility is manifested by the breaking of the trees at the point
of the union particularly when they have been growing for some years (apricot on Prunus
grafts, pear on quince grafts).
This review provides a summary of the new advances in studies on the mechanism of
graft compatibility focused on the early responses of grafting and how these studies can be
correlated with the changes observed in some Prunus combinations at early stages.
incompatibility in woody plants (Mosse, 1962; Errea et al., 1994a,b). In these cases, an
abnormal process of neocambium differentiation leads to a cambial involution and a lack
of differentiation into new vascular elements, as has been pointed out for pear and
quince grafts (Ermel et al., 1999) and apricot on Prunus grafts (Errea et al., 1994b).
While it appears that the formation of functional vascular connections is essential for
successful grafts in herbaceous plants, in woody plants incompatible grafts can grow for
several years without any external indication of incompatibility, denoting the presence
of functional vascular connections in incompatible grafts (Mosse, 1962; Hartmann et al.,
1997).
Studies on phloem regeneration across the graft interface have been the subject of
some herbaceous investigations (Stoddard and Mc Cully, 1979; Tiedemann, 1989;
Kollmann and Glockmann, 1990; Golecki et al., 1998). In the compatible system
Lycopersicon esculentum (L) on Solanum tuberosum (S) as well as in the autografts
(Scho ning and Kollmann, 1995, 1997) 14C-labelling techniques revealed that
assimilated transport occurs from the apical callus of the scion, to the basal callus
of the stock. As was expected, a close interrelation between transport and phloem
restitution in the graft union could be demonstrated. On the contrary, in incompatible
system Vicia faba (V) on Helianthus annuus (H), an increase in 14C-transport to the
stock during all stages of graft union development did not occur and no correlation was
found between phloem regeneration and assimilation transport across the graft interface
in this less compatible combination. 56 carboxifluorescein (CF) translocation
experiments confirm non-functional phloem connections in V/H-heterograft (Scho ning
and Kollmann, 1997).
However, in apricot (Prunus armeniaca) on plum grafts, the formation of new vascular
connections occurs in both compatible and incompatible combinations (Errea et al.,
1994b). The transport of disodium fluorescein across the graft union confirms the
communication and functionality of these connections since fluorescence can be seen in
both partners of the graft. In these combinations, the difference between compatible and
incompatible grafts lies in the presence of a portion of the callus tissue in incompatible
grafts that cannot differentiate into cambium and vascular tissue, resulting in the existence
of wide areas at the union similar to undifferentiated callus cells. This lack of cambial
activity in some areas of the graft union could affect the activity of the new xylem and
phloem formed, causing discontinuities in the cambium and the formation of a
parenchymatous line interrupting the vascular connection (Hartmann et al., 2002),
producing a mechanically weak union.
The majority of studies on herbaceous grafts have been performed with Cucurbits due
to their distinct phloem anatomy and plentiful vascular exudation. As it is known, during
the ontogeny of the sieve element/companion cell complex, synthesis of proteins
coincides with vascular development. Evidence suggests that in the graftage of Cucumis
and Cucurbita, changes in protein banding may be due to polypeptides migrating
symplastically across the graft union via the connecting phloem and not from the
grafting procedure (Tiedemann and Carsens-Behrens, 1994). The urgent need to
reconnect disrupted vascular bundles should result in the expression of essential
proteins, possibly at reduced concentrations owing to abbreviated developmental periods
(Schulz, 1990). The question opened so far is whether in some Prunus grafts, where good
role and effects on incompatibility is still not clear (Quesada and Macheix, 1984; Deloire
and Hebant, 1982). Schmid and Feucht (1985) had studied proteins, peroxidases in the
phloem and acid phosphatases in Prunus avium/Prunus cerasus grafting confirming that is
possible to define a good union not only by morphological characters but also by
biochemical methods. Based on biochemical assays, Gulen et al. (2002) concluded that the
absence or presence of peroxidase isozymes profile in the pear-quince graft combination is
an experimental approach to predict incompatibility reaction. More recently, investigations
suggest that increased peroxidase and catalase activities might be involved in graft
development in tomato plants (Fernandez-Garcia et al., 2004).
With regard to grafting process in fruit trees more studies should be performed to obtain
a deeper knowledge about the mechanisms that take place during the graft incompatibility
reactions, which will allow us to make an early rootstock selection, before we observe any
external incompatibility symptoms.
5. Conclusions
The developmental process of the graft union has attracted considerable attention and
much information about it has been gathered. Several investigators have commented on
structural events that may be responsible for the development of cohesion between the stock
and scion during graft ontogeny. These reports study cytological events occurring in response
to grafting with special reference to the possibility that the phenomena of cellular recognition
could be involved in the development of functional vascular connections, since the basis of an
incompatibility response could be determined by local interactions between the opposing
cells of the graft union itself. Contrary to this hypothesis, the cohesion of graft partners has
been explained as resulting from the deposition and subsequent polymerization of cell wall
material that occur in response to the wounding inherent to graft establishment, which is not
related to the compatibility reaction. Despite the fact that callus formation can be considered
as a common wound healing response in plants, recent advances in studying the
plasmodesmata as the highly dynamic structures that offer a pathway for symplastic cell
communication, open the door to its important role in cell recognition, and compatibility and
incompatibility response. Also other lines of research using grafting techniques, revealed the
existence of soluble proteins in phloem exudates that have been developmentally related to
defined stages of phloem differentiation; more experimental proofs will be necessary in order
to fully understand their involvement in phloem function. As the mechanisms involved in all
these processes have been related to herbaceous graft combinations, future studies should
include a much wider range of similarities on woody responses to reach a better
understanding of the mechanism of incompatibility in these graft combinations.
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