Preface

Grafting is essential to fruit tree propagation. Using the scion wood you essentially create a clone
of your parent tree and so keep the variety true to form. Fruit varieties cannot be propagated any
other way.

To produce a tree of a known variety, you need to graft from material of that variety. To do this
with fruit trees you take scions (essentially new twigs) or buds, and graft them on to a rootstock.
You then have the roots of one tree, and the branches, leaves and fruits of another; your chosen
fruit variety. In fact, most fruit trees are cultivated like this as fruit tree varieties don’t grow true
from pips. If you plant the pip of a Bramley’s seedling apple, the resultant tree will be an apple,
but not a Bramley. To get a Bramley, you need to cut off a scion or a vegetative bud, and graft it
on to a rootstock. This essentially clones the tree, growing one with the exact genetic makeup so
preserving the variety true to form. If you think about it, this means all Bramley trees have
exactly the same genetic makeup as that first original tree.

It might sound like a daunting process but grafting can actually be quite straight forward. As
long as you follow a few simple rules, you should find that you make successful grafts from the
start. The two most crucial considerations for successful grafting are timing, and cambium
contact. These will vary depending on the types of graft you are attempting.

Grafting provides a simple way to generate chimeric plants with regions of different genotypes,
and thus to assess the cell autonomy of gene action. The technique of grafting has been widely
used in other species, but in Arabidopsis, its small size makes the process rather more
complicated. However, there are now several well-established grafting procedures available,
which we described here, and their use has already contributed greatly to understanding of such
processes as shoot branching control, flowering, and disease resistance.

Since the origins of agriculture, the progressive domestication of food crops has been intimately
related to a series of innovations in plant propagation. In the Fertile Crescent, 10,000 to 12,000
years ago, nomadic peoples subsisted in part by collecting the seeds of autogamous (self-
pollinating) wild grasses (emmer and einkorn wheat, barley) and pulses (lentils, chickpeas, peas).
Unintentional selection for traits useful to humans, such as nonshattering, uniform ripening, and

1
other characteristics known as the domestication syndrome, progressed relatively rapidly. This
resulted in some measure of food security, which contributed to social stability and the decline of
nomadism. Although fruits, nuts, and other tree-related foods and fibers were an important part
of the diet, it was only several thousands. of years later that domestication of woody plants
began. Unlike the grains and pulses domesticated much earlier, most useful woody species are
highly heterozygous and do not come true to type from seed, which precluded rapid genetic
improvement by seedling selection. What made tree domestication possible was the mastery of
cloning via asexual propagation. Initially this involved cloning of genetically superior
individuals of species that rooted easily from cuttings or layering or propagated by offshoots.
The modification and adoption by early agriculturists of these techniques allowed for the
domestication of fig, grape, pomegranate, and olive in the third or fourth millennium; all of
which root easily from cuttings, and date palm, which was propagated by division of offshoots.
The domestication of woody species that do not root easily from cuttings, such as apples, pears
and plums, did not come until the discovery of grafting, at least several thousand years later,
about the beginning of the first millennium BCE. Thus, grafting is a pivotal technology in the
history of temperate fruits and probably influenced their movement from Central Asia to Europe.

2
 Chapter No.1. Introduction

Grafting is among the most expensive propagation techniques, surpassing even micro
propagation. Since the beginning of civilization, fruit and nut trees have been grafted because of
the difficulty in propagating by cuttings, and the superiority and high value of the grafted crop.
Budding, which is a form of grafting, is three times more costly than cuttings and fourteen times
more expensive than seedling propagation. The horticulture and forestry industries have sought
to develop clonal propagation systems that avoid labor-intensive graftage. Yet, traditional and
highly efficient grafting and budding systems are essential for the propagation of many woody
plant species. New markets continue to require grafted and budded plants for improved plant
quality, fruit yield, superior forms, and better adaptation to greater ecological ranges. The
propagator benefits via new markets, while the consumer gains a greater variety of better adapted
landscape plants. The acid-loving blueberry can be produced in more basic pH soils when
grafted to pH-tolerant rootstock. With the greater reliance on integrated pest management and
reduced availability of pesticides and soil fumigants, disease-tolerant rootstocks are playing a
greater role not only with woody perennial fruit crops and ornamentals, but also with grafted
vegetable crops.

Grafting is the art of joining two species of living plant tissue together in such a manner that they
will unite and subsequently grow and develop as one composite plant.

Budding is a form of grafting. However, the scion is reduced in size and usually contains only
one bud. An exception to this is patch budding of pecan, where secondary and tertiary buds are
adjacent at the same node to the primary bud.

The scion becomes the new shoot system of the graft. It is composed of a short piece of detached
shoot containing several dormant buds, which, when united with the rootstock, comprises the
upper portion of the graft. The stem, or branches, or both, of the grafted plant will grow from the
scion. The scion should be of the desired cultivar and free from disease.

The rootstock is the lower portion of the graft, which develops into the root system of the grafted
plant. It may be a seedling, a rooted cutting, a layered or micro propagated plant. If the grafting
is done high in a tree, as in top working, the rootstock may consist of the roots, trunk, and
scaffold branches.
3
The interstock is a piece of stem inserted by means of two graft unions between the scion and the
rootstock. Interstocks are used to avoid incompatibility between the rootstock and scion, to
produce special tree forms, to control disease (e.g., fire-blight resistance), or to take advantage of
their growth-controlling properties.Vascular cambium is a thin tissue located between the bark
and the wood. Its cells are meristematic that is, they are capable of dividing and forming new
cells.

For a successful graft union, it is essential that the cambium of the scion be placed in close
contact with the cambium of the rootstock.

Callus is a term applied to the mass of parenchyma cells that develop from and around wounded
plant tissues. It occurs at the junction of a graft union, arising from the living cells of both the
scion and rootstock. The production and interlocking of these parenchyma (or callus) cells
constitute one of the important steps in callus bridge formation between the scion and rootstock
in a successful graft.

4
 Chapter No.2. Review of Literature

Plant grafting is an important horticulture technique used to produce a new plant after joining
rootstock and scion. This is one of the most used techniques by horticulturists to enhance the
quality and production of various crops. Grafting helps in improving the health of plants, their
yield, and the quality of plant products, along with the enhancement of their postharvest life. The
main process responsible for successful production of grafted plants is the connection of vascular
tissues. This step determines the success rate of grafts and hence needs to be studied in detail.
There are many factors that regulate the connection of scion and stock, and plant hormones are
of special interest for researchers in the recent times. These phytohormones act as signaling
molecules and have the capability of translocation across the graft union. Plant hormones, mainly
auxins, cytokinins, and gibberellins, play a major role in the regulation of various key
physiological processes occurring at the grafting site. In the current review, we discuss the
molecular mechanisms of graft development and the phytohormone-mediated regulation of the
growth and development of graft union (Anket Sharma and Bingsong Zheng 2019).

Hormonal signals, auxin in particular, are believed to play an important role in the wound
healing and vascular regeneration within the graft union zone. Incomplete and convoluted
vascular connections impede the vital upward and downward whole plant transfer routes.
Longdistance protein, mRNA and small RNA graft-transmissible signals currently emerge as
novel mechanisms which regulate nutritional and developmental root/top relations and may play
a pivotal role in grafting physiology. Grafting also has significant pathogenic projections. On one
hand, stock to scion mechanical contact enables the spread of diseases, even without a complete
graft union. But, on the other hand, grafting onto resistant rootstocks serves as a principal tool in
the management of fruit tree plagues and vegetable soil-borne diseases.The ‘graft hybrid’
historic controversy has not yet been resolved. Recent evidence suggests that epigenetic
modification of DNA-methylation patterns may account for certain graft-transformation
phenomena. Root grafting is a wide spread natural phenomenon; both intraspecific and
interspecific root grafts have been recorded. Root grafts have an evolutionary role in the survival
of storm-hit forest stands as well as in the spread of devastating diseases. A more fundamental
evolutionary role is hinted by recent findings that demonstrate plastid and nuclear genome
transfer between distinct Nicotiana species in the graft union zone, within a tissue culture system.

5
This has led to the formation of alloploid cells that, under laboratory conditions, gave rise to a
novel, alloploid Nicotiana species, indicating that natural grafts may play a role in plant
speciation, under certain circumstances ( Eliezer E. Goldschmidt 2014)

Grafting is a widely utilized agronomical technique to improve yield, disease resistance, and
quality of fruit and vegetables. This work aims to assess the effect of grafting and fruit ripening
on the production, physico-chemical characteristics, and nutritional quality of fruit from Spanish
local pepper landraces. Landraces “Cuerno,” “Sueca,” and “Valencia” were used as scions, and
“NIBER” as the rootstock. Two ripening stages of the fruits were sampled: green and red.
Grafting improved the yield and marketable quality and did not negatively influence the physico-
chemical and nutritional characteristics of the fruit. It was noteworthy that the bioactive
compound contents and antioxidant capacity were more related to maturity stage and genotype,
and red fruit had a higher antioxidant capacity than green fruit. However, in all the scions,
grafting significantly enhanced lycopene content in both red and green fruit. Another important
effect of grafting was the volatile compound composition evidenced by discriminant analyses,
which was characterized for the first time in the fruit of these landraces. The rootstock and scion
combination could be a way to improve not only the production, but also the fruit quality of
peppers( Ramón Gisbert-Mullor et.al.2020).

For millennia, people have cut and joined different plant tissues together through a process
known as grafting. By creating a chimeric organism, desirable properties from two plants
combine to enhance disease resistance, abiotic stress tolerance, vigour or facilitate the asexual
propagation of plants. In addition, grafting has been extremely informative in science for
studying and identifying the long-distance movement of molecules. Despite its increasing use in
horticulture and science, how plants undertake the process of grafting remains elusive. Here, we
discuss specifically the role of eight major plant hormones during the wound healing and
vascular formation process, two phenomena involved in grafting. We furthermore present the
roles of these hormones during graft formation and highlight knowledge gaps and future areas of
interest for the field of grafting biology (Amrit K. Nanda and Charles W. Melnyk . 2017).

Grafting has been widely used to improve horticultural traits. It has also served increasingly as a
tool to investigate the long-distance transport of molecules that is an essential part for key

6
biological processes. Many studies have revealed the molecular mechanisms of graft-induced
phenotypic variation in anatomy, morphology and production ( Jing Wang. et.al 2016).

The majority of homografts are compatible, with the exception of monocots. Since the wound
required for grafting disrupts the plant vascular system (Asahina & Satoh, 2015)

Although grafting is widely used in the agriculture of fruit-bearing crops, little is known about
graft union formation in particular when two different species are grafted together. It is
fascinating that two different plant species brought together can develop harmoniously as one
organism for many decades. The objective of this study was to determine whether grafting two
different grapevine genotypes alters gene expression at the graft interface in comparison to the
presumably wound-like gene expression changes induced in auto grafts. Gene expression at the
graft interface was studied 3, 7, 14, and 28 d after grafting in hetero- and auto grafts of grapevine
(Vitis spp.). Genes differentially expressed between the hetero- and auto grafts during graft
union formation were identified. These genes were clustered according to their expression profile
over the time course. Map Man and Gene Ontology enrichment analysis revealed the coordinated
upregulation of genes from numerous functional categories related to stress responses in the
hetero- compared to the auto grafts. This indicates that heterografting with nonself rootstocks
upregulates stress responses at the graft interface, potentially suggesting that the cells of the graft
interface can detect the presence of a nonself grafting partner (S.J.Cookson et.al 2014).

Grafting is particularly important to the cultivation of perennial crops such as grapevine (Vitis
vinifera) because rootstocks can provide resistance to soil-borne pests and diseases as well as
improve tolerance to some abiotic stresses. Successful grafting is a complex biochemical and
structural process beginning with the adhesion of the two grafted partners, followed by callus
formation and the establishment of a functional vascular system. At the molecular level, the
sequence of events underlying graft union formation remains largely uncharacterized. The
present study investigates the transcriptome of grapevine rootstock and graft interface tissues
sampled 3 d and 28 d after grafting of over-wintering stems in the spring. Many genes were
differentially expressed over time, from 3 d to 28 d after grafting, which could be related to the
activation of stem growth and metabolic activity in the spring. This hypothesis is supported by
the up-regulation of many genes associated with cell wall synthesis, and phloem and xylem
development. Generally, there was an up-regulation of gene expression in the graft interface

7
tissue compared with the rootstock, particularly genes involved in cell wall synthesis, secondary
metabolism, and signalling. Although there was overlap between the genes differentially
expressed over time (from 3 d to 28 d after grafting) with the gene differentially expressed
between the rootstock and the graft interface, numerous graft interface-specific genes were
identified (Sarah Jane Cookson et. al 2013) .

Laccase‐mediated grafting on lignocelluloses has gained considerable attention as an

environmentally benign method to covalently modify wood, paper and cork. In recent decades
this technique has also been employed to modify fibers with a polysaccharide backbone, such as
cellulose or chitosan, to infer colouration, antimicrobial activity or antioxidant activity to the
material. The scope of this approach has been further widened by researchers, who apply
mediators or high redox potential laccases and those that modify synthetic polymers and
proteins. In all cases, the methodology relies on one‐ or two‐ electron oxidation of the surface
functional groups or of the graftable molecule in solution. However, similar results can very
often be achieved through simple deposition, even after extensive washing. This unintended
absorption of the active substance could have an adverse effect on the durability of the applied
coating. Differentiating between actual covalent binding and adsorption is therefore essential, but
proves to be challenging. This review not only covers excellent research on the topic of laccase‐
mediated grafting over the last five to ten years, but also provides a critical comparison to
highlight either the lack or presence of compelling evidence for covalent grafting(Sjoerd
Slagman et. al 2017).

Grafting is an excellent tool to study long-range signaling processes in plants. And the grafting
between species will help elucidate the molecular mechanisms underlying contrasting differences
between different species. Arabidopsis thaliana is a salt-sensitive model glycophyte and Eutrema
salsugineum (previously Thellungiella salsuginea, salt cress) is a salt-tolerant model halophyte.

Successful grafting of these two model plants will help further study the physiological and
molecular mechanisms underlying salt tolerance in plants. The aim of this study was to
demonstrate two sterile micro-grafting methods for Arabidopsis and salt cress (Yan Li et. al
2019).

8
 The extreme phenotype of zinc (Zn) hyperaccumulation, which is found in several
Brassicaceae species, is determined by mechanisms that promote elevated Zn tolerance and high
Zn accumulation in shoots( Marcelo de A Guimaraes et. al 2009 ).

Grafting has long been an important technique in agriculture. Nowadays, grafting is a widely
used technique also to study systemic long-distance signaling in plants. Plants respond to their
surrounding environment, and at that time many aspects of their physiology are regulated
systemically; these start from local input signals and are followed by the transmission of
information to the rest of the plant. For example, soil nutrient conditions, light/photoperiod, and
biotic and abiotic stresses act plants heterogeneously, and plants perceive such information in
specic plant tissues or organs. Such environmental cues are crucial determinants of plant growth
and development, and plants drastically change their morphology and physiology to adapt to
various events in their life. Hitherto, intensive studies have been conducted to understand
systemic signaling in plants, and grafting techniques have permitted advances in this eld. The
breakthrough technique of micrografting in Arabidopsis thaliana was established in 2002 and led
to the development of molecular genetics tools in this eld. Thereafter, various phenomena of
systemic signaling have been identied at the molecular level, including nutrient xation, owering,
circadian clock and defense against pathogens. The signicance of grafting is that it can clarify the
transmission of the stimulus and molecules. At present, many micro- and macromolecules have
been identied as mobile signals, which are transported through plant vascular tissues to co-
ordinate their physiology and development. In this review, we introduce the various grafting
techniques that have been developed, we report on the recent advances in the field of plant
systemic signaling where grafting techniques have been applied and provide insights for the
future ( Hiroki Tsutsui &Michitaka Notaguch 2017) .

Even today grafting is applied to propagate various fruit trees, such as apples, pears, citrus,
persimmons and grapes. The purpose of grafting is varied; the most common use is the clonal
vegetative propagation of heterogenic trees to harbor desirable traits. In addition, grafting is used
to control size/growth, shorten juvenility and gain biotic and abiotic stress tolerance. The
methodology has been improved to enhance grafting success and achieve higher quality traits.
While many methods are reported, there is a basic common principle, i.e. cutting and assembling
the plant parts that have good tissue tness and unifying them to grow on the grafted plants as a

9
singular plant. Today, grafting is also applied to vegetable cultivation in Asia, Europe, the
Middle East, Northern Africa and Central America. For example, Cucurbitaceae spp. including
cucumbers, watermelons and pumpkins, and Solanaceae spp. including tomatoes (Solanum
lycopersicon) have been grafted to enhance their resistance to soil-borne diseases/nematodes,
increase yields and enhance fruit quality (Kubota et al. 2008).

Several endings of light transmitting information to plants have been recently reported through
the use of grafting. A bZIP transcription factor, elongated hypocotyl5 (HY5), regulates plant
growth in response to light. Recently, grafting experiments using hy5 loss-of-function mutants
uncovered the systemic action of HY5 to regulate root growth and nitrate uptake depending on
light illumination of shoots. Its transport was detected by grafting, using the HY5–green
uorescent protein (GFP) fusion construct, and the authors concluded that C assimilation in shoots
and N absorption in roots were co-ordinated through the transport of HY5 from shoot to root,
which promoted root nitrate uptake by activating the expression of a nitrate transporter gene
(Chen et al. 2016).

The photoperiod is an important environmental cue that provides essential information for
plants, allowing them to recognize seasons in broad areas on the planet. The information is
mainly perceived in mature leaves and transmitted throughout the plant. Plant photoperiodic
responses are varied and may include owering in some species, tuberization in potatoes and bud
set in trees, and much progress has been made toward understanding the molecular mechanisms
(Jackson 2009)

ABA. Shoot-to-root translocation of ABA was reported to promote root growth in tomato and
eld pea under well-watered conditions (McAdam et al. 2016).

Jasmonic acid (JA) is involved in wound response and is triggered by wounding such as insect
damage and mechanical stimulus. The wound signal spreads systemically to induce the
expression of defense genes in unwounded tissues (Stratmann 2003).

The grafting of fruits and vegetables influences fruit quality. The aim of the present work was
to assess the effect of the rootstock and the scion on the antioxidant activity and the content in
vitamin C, total phenols, lycopene and β-carotene of bell pepper. The cultivars Fascinato and
Jeanette were used as scion and Terrano was used as rootstock. Four harvests in the production

10
cycle of the vegetable were analyzed in a cultivation system under shading nets. The results
indicate statistical differences in the content of these bioactive compounds between the varieties,
between grafting and not grafting and between sampling dates (p ≤ 0.05). The vitamin C content,
β-carotene, and antioxidant capacity proved significantly higher in Fascinato than in Janette. On
average, grafting increased β-carotene and vitamin C concentrations and improved the
antioxidant capacity, but had no influence on the total phenol or lycopene contents. It is
concluded that grafting to the rootstock Terrano improves the nutritional quality of the fruit
produced in both varieties of bell pepper studied (Celia Chávez-Mendoza et. al 2013).

The majority of homografts are compatible, with the exception of monocots. Since the wound
required for grafting disrupts the plant vascular system (Asahina & Satoh, 2015).

When the cambium of the scion joins fully with that of the rootstock, intact cells divide and
proliferate into calli, which eventually differentiate into vasculature and plasmodesmata forms
(Melnyk & Meyerowitz 2015).

After cell walls fuse in the graft union, plasmodesmata stretch in small groups over the spaces of
the inner cell wall, interconnecting the protoplasts of contiguous cells (Kollmann & Glockmann,
1985).

The plasmodesmata provide tunnels for small molecules and even selectively permit the
movement of macromolecules, such as proteins and nucleic acids. Additionally, vascular
reconstruction at the graft union enables macromolecules to be transported (Harada, 2010).

In recent years, increasing effort has been made to determine how macromolecules are
transferred between scions and rootstocks in grafting plants to reveal the mechanisms that
control graft-induced changes in plant traits (Paultre et al 2016).

Grafting commonly influences the phenotype of the grafted plants (Warschefsky et al 2015).

Grafting is widely used to improve resistance to pests and diseases. For instance, grafting can
alleviate the development of post-harvest diseases in Hass avocado fruit (Willingham et al2001).

11
 Chapter No.3. Factors for Successful Graft

Compatibility of scion and stock: Grafting involves the joining of vascular tissues between the
scion and rootstock, plants lacking vascular cambium, such as monocots, cannot normally be
grafted. As a general rule, the closer two plants are genetically, the more likely the graft union
will form. Genetically identical clones and intra-species plants have a high success rate for
grafting. Grafting between species of the same genus is sometimes successful. Grafting has a low
success rate when performed with plants in the same family but in different genera and grafting
between different families is rare.

Cambium alignment and pressure: The vascular cambium of the scion and stock should be
tightly pressed together and oriented in the direction of normal growth. Proper alignment and
pressure encourages the tissues to join quickly, allowing nutrients and water to transfer from the
stock root to the scion.

Completed during appropriate stage of plant: The grafting is completed at a time when the
scion and stock are capable of producing callus and other wound-response tissues. Generally,
grafting is performed when the scion is dormant, as premature budding can drain the grafting site
of moisture before the grafting union is properly established. Temperature greatly affects the
physiological stage of plants. If the temperature is too warm, premature budding may result.

Proper care of graft site: After grafting, it is important to nurse the grafted plant back to
health for a period of time. Various grafting tapes and waxes are used to protect the scion and
stock from excessive water loss. Furthermore, depending on the type of graft, twine or string is
used to add structural support to the grafting site. Sometimes it is necessary to prune the site, as
the rootstock may produce shoots that inhibit the growth of the scion.

12
 Chapter No. 4. Methods

Main methods of grafting are as follows :

Approach: Approach grafting or inarching is used to join together plants that are otherwise
difficult to join. The plants are grown close together, and then joined so that each plant has roots
below and growth above the point of union. Both scion and stock retain their respective parents
that may or may not be removed after joining. Also used in pleaching. The graft can be
successfully accomplished any time of year.

Figure No.1 : Approach Grafting

Bud

Bud grafting (also called chip budding) uses a bud instead of a twig. Grafting roses is the most
common example of bud grafting. In this method a bud is removed from the parent plant, and the

13
base of the bud is inserted beneath the bark of the stem of the stock plant from which the rest of
the shoot has been cut. Any extra bud that starts growing from the stem of the stock plant is
removed. Examples: roses and fruit trees like peaches.

Figure No.2: Bud Grafting

Cleft

In cleft grafting a small cut is made in the stock and then the pointed end of the scion is inserted
in the stock. This is best done in the early spring and is useful for joining a thin scion about 1 cm
diameter to a thicker branch or stock. It is best if the former has 3–5 buds and the latter is 2–7 cm
in diameter. The branch or stock should be split carefully down the middle to form a cleft about
3 cm deep. If it is a branch that is not vertical then the cleft should be cut horizontally. The end
of the scion should be cut cleanly to a long shallow wedge, preferably with a single cut for each
wedge surface, and not whittled. A third cut may be made across the end of the wedge to make it
straight across. Slide the wedge into the cleft so that it is at the edge of the stock and the centre of

14
the wedge faces are against the cambium layer between the bark and the wood. It is preferable if
a second scion is inserted in a similar way into the other side of the cleft. This helps to seal off
the cleft. Tape around the top of the stock to hold the scion in place and cover with grafting wax
or sealing compound. This stops the cambium layers from drying out and also prevents the
ingress of water into the cleft.

Figure No.3: Cleft Grafting

Whip

In whip grafting the scion and the stock are cut slanting and then joined. The grafted point is then
bound with tape and covered with a soft sealant to prevent dehydration and infection by germs.
The common variation is a whip and tongue graft, which is considered the most difficult to
master but has the highest rate of success as it offers the most cambium contact between the
scion and the stock. It is the most common graft used in preparing commercial fruit trees. It is

15
generally used with stock less than 1.25 cm diameter, with the ideal diameter closer to 1 cm and
the scion should be of roughly the same diameter as the stock.

The stock is cut through on one side only at a shallow angle with a sharp knife. (If the stock is a
branch and not the main trunk of the rootstock then the cut surface should face outward from the
centre of the tree.) The scion is similarly sliced through at an equal angle starting just below a
bud, so that the bud is at the top of the cut and on the other side than the cut face. In the whip and
tongue variation, a notch is cut downwards into the sliced face of the stock and a similar cut
upwards into the face of the scion cut. These acts as the tongues and it require some skill to make
the cuts so that the scion and the stock marry up neatly. The elongated "Z" shape adds strength,
removing the need for a companion rod in the first season. The joint is then taped around and
treated with tree-sealing compound or grafting wax. A whip graft without a tongue is less stable
and may need added support.

Figure No.4: Whip Grafting

16
Stub

Stub grafting is a technique that requires less stock than cleft grafting, and retains the shape of a
tree. Also scions are generally of 6–8 buds in this process. An incision is made into the branch 1
cm long, then the scion is wedged and forced into the branch. The scion should be at an angle of
at most 35° to the mother plant so that the crotch remains strong. The graft is covered with
grafting compound. After the graft has taken, the branch is removed and treated a few
centimeters above the graft, to be fully removed when the graft is strong.

Figure No.5: Stub Grafting

Four-Flap

The four-flap graft (also called banana graft) is commonly used for pecans, and first became
popular with this species in Oklahoma in 1975. It is heralded for maximum cambium overlap,
but is a complex graft. It requires similarly sized diameters for the rootstock and scion. The bark
of the rootstock is sliced and peeled back in four flaps and the hardwood is removed, looking
somewhat like a peeled banana. It is a difficult graft to learn.

17
 Figure No.6: Four – Flap Graft

Awl

Awl grafting takes the least resources and the least time. It is best done by an experienced
grafter, as it is possible to accidentally drive the tool too far into the stock, reducing the scion's
chance of survival. Awl grafting can be done by using a screwdriver to make a slit in the bark,
not penetrating the cambium layer completely. Then inset the wedged scion into the incision.

Figure No.7: Awl Grafting

18
Veneer grafting

Veneer grafting, or inlay grafting, is a method used for stock larger than 3 cm in diameter. The
scion is recommended to be about as thick as a pencil. Clefts are made of the same size as the
scion on the side of the branch, not on top. The scion end is shaped as a wedge, inserted, and
wrapped with tape to the scaffolding branches to give it more strength.

Figure No.8: Veneer grafting

Rind

Rind grafting involves grafting a small scion into the end of a thick stock. The thick stock is
sawn off, and a 4 cm long bark-deep cut is made parallel to the stock, from the sawn-off end
down, and the bark is separated from the wood on one or both sides. The scion is shaped as a
wedge, exposing cambium on both sides, and is pushed in under the back of the stock, with a flat
side against the wood.

19
 Figure No.9: Rind Grafting

Natural Grafting

Tree branches and more often roots of the same species will sometimes naturally graft; this is
called inosculation. The bark of the tree may be stripped away when the roots make physical
contact with each other, exposing the vascular cambium and allowing the roots to graft together.

Natural grafting is rarely seen in herbaceous plants as those types of plants generally have short-
lived roots with little to no secondary growth in the vascular cambium.

20
 (A) (B)

Figure No.10: A&B Natural Grafting

Graft Chimera

Graft chimera can occur where the tissues of the stock continue to grow within the scion. Such a
plant can produce flowers and foliage typical of both plants as well as shoots intermediate
between the two.

Figure No.11: Graft chimera in cactus

21
 Chapter No.5. Materials

Cutting tools: It is good procedure to keep the cutting tool sharp to minimize tissue damage and
clean from dirt and other substances to avoid the spread of disease. A good knife for general
grafting should have a blade and handle length of about 3 inches and 4 inches respectively.
Specialized knives for grafting include bud-grafting knives, surgical knives, and pruning knives.
Cleavers, chisels, and saws are utilized when the stock is too large to be cut otherwise.

Figure No.12: Cutting tools

22
Disinfecting materials: Treating the cutting tools with disinfectants ensures the grafting site is
clear of pathogens. A common sterilizing agent is absolute alcohol.

Figure No.13: Disinfecting materials

Graft seals: Keeps the grafting site hydrated. Good seals should be tight enough to retain
moisture while, at the same time, loose enough to accommodate plant growth. Includes
specialized types of clay, wax, petroleum jelly, and adhesive tape.

Figure No.14: Graft seals (Home made)

23
Tying and support materials: Adds support and pressure to the grafting site to hold the stock
and scion together before the tissues join, which is especially important in herbaceous grafting.
The employed material is often dampened before use to help protect the site from desiccation.
Support equipment includes strips made from various substances, twine, nails, and splints.

Figure No.15: Tying and support materials

Grafting machines: Because grafting can take a lot of time and skill, grafting machines have
been created. Automation is particularly popular for seedling grafting in countries such as Japan
and Korea where farming land is both limited and used intensively. Certain machines can graft
800 seedlings per hour.

24
Figure No.16: Grafting machines

25
 Chapter No.6. Result &Discussion

Grafting is a horticulture technique where the living detached portion of the plant is used to regenerate
the growth .One plant is selected for its roots and this is called the stock or rootstock .The other plant
is selected for its stems, leaves, flowers, or fruits and the scion. The joining of scion and stalk is
known as inosculation.
Grafting involves mixing of traits of two varieties without resorting to hybridization as because stock
and scion belong to two different plant species. The vegetative part is developed by the scion so fruit,
flowering, branching, and leaves pattern depends on scion while the root pattern is influenced by the
genome of the stock.
The best time for grafting is in the spring just as growth starts. When necessary, grafting can start
several weeks before growth is expected and can continue a few weeks after growth has started,
if you have dormant scion wood in storage and if weather is not exceptionally warm.
In a successful graft you should see the scion plump up. If it still looks somewhat flaccid it's
likely it didn't take. Others with more experience will hopefully chime in.Cleft and
contact graft resulted more success than side grafting. The maximum 9.67 (out of 10) graft
success with 86.67% establishment was obtained in April from cleft grafting but the maximum
establishment 90% was obtained from contact grafting in May without any death of
separated grafts.

(A) (B)

Figure No.17: (A) Red hibiscus grafted with pink hibiscus and (B) Dark red hibiscus grafted with yellow hibiscus

26
 Chapter No.7. Conclusion

The technology of grafting has advanced tremendously; the long-term survival of grafted,
composite plants is still somewhat unpredictable. The complexity of stock/scion interactions
attains a new dimension when pathogenic agents enter the scenery. More enigmatic are the graft
hybrid conundrum and the broader evolutionary significance of grafting. The extensive
horticultural grafting research has not usually addressed these basic questions. The recently
adopted Arabidopsis model has opened the way for meticulous, in-depth grafting research. The
currently available molecular tools are expected to advance our understanding and eventually
resolve the long standing grafting mysteries.

27
 Chapter No.8. Reference
1. Amrit K. Nanda,Charles W. Melnyk12017.The role of plant hormones
during grafting. doi.org/10.1007/s10265-017-0994-5
2. Ahlswede, J. 1985. Twig grafting of macadamia. Comb. Proc. Intl. Plant Prop. Soc.
34:211–14.
3. Alley, C. J. 1957. Mechanized grape grafting. Calif. Agr. 11:3,12.
4. Alley, C. J. 1970. Can grafting be mechanized? Comb. Proc. Intl. Plant Prop. Soc.
20:244–48.
5. Anonymous. 1968. Pre-cut scions. Agr. Res. 17:11.
6. Asahina M, Satoh S. 2015. Molecular and physiological mechanisms regulating tissue
reunion in incised plant tissues. Journal of Plant Research 128: 381–388
7. Beineke, W. F, 1978. Parafilm: A new way to wrap grafts. HortScience. 13:284.
8. Bhar, D. S., R. J. Hilton, and G. C. Ashton. 1966. Effect of time of cutting and
storage treatment on growth and vigor of scions of Malus pumila cv. McIntosh. Can.
J. Plant Sci. 46:69–72.
9. Brase, K. D. 1951. The nurse-root graft, an aid in rootstock research. Farm Res.
17:16.
10. Burger, D. W. 1985. Micrografting: A tool for the plant propagator. Comb. Proc. Intl.
Plant Prop. Soc. 34:244–48.
11. Carpenter, E. L. 1989. How records can improve grafting. Comb. Proc. Intl. Plant
Prop. Soc. 39:413–15. 10. Cummins, J. N. 1973. Systems for producing multiple-
stock fruit trees in the nursery. Plant Propagator. 19:7–9.
12. "Citrus Budwood Program" (http://www.freshfromflorida.com/Divisions-
Offices/Plant-Industry/Agricult ure-Industry/Citrus-Health-Response-Program/Citrus-
Budwood-Program). Florida Department of Agriculture and Consumer Services.
Retrieved 13 September 2017.
13. "Citrus Budwood Cerfication Program" (http://kcc-weslaco.tamu.edu/budwood.html).
Texas A&M University - Kingsville Citrus Center. September 29, 2015. Retrieved 13
September 2017.

28
14. Cohen, R., Y. Burger, C. Horev, A. Porat, and M. Edelstein. 2005. Performance of
Galia type melons grafted onto Cucurbita rootstock in Monosporascus cannonballus-
infested and non-infested soils. Ann. Appl.Biol. 146:381–87.
15. Cookson S.J., Moreno M.J. Clemente , Hevin C., Mendome L.Z. Nyamba, Delrot
S., Trossat-Magnin C., Ollat N,2013.Graft union formation in grapevine induces
transcriptional changes related to cell wall modification, wounding, hormone
signalling, and secondary metabolism . doi: 10.1093/jxb/ert144.
16. Chávez-Mendoza C., Sánchez E , Carvajal-Millán E., Marquez E.M., Guevara-
Aguilar A. 2013. Characterization of the nutraceutical quality and antioxidant activity
in bell pepper in response to grafting. doi: 10.3390/molecules181215689
17. Chen X., Yao Q., Gao X., Jiang C., Harberd N.P., Fu X. (2016) Shoot-to-root mobile
transcription factor HY5 coordinates plant carbon and nitrogen acquisition. Curr.
Biol. 26: 640–646
18. Davis, A. R., P. Perkins-Veazie, R. Hassell, S. R. King, and X. Zhang. 2008a.
Grafting effects on vegetable quality. HortScience 43:1670–2.
19. Davis, A. R., P. Perkins-Veazie, Y. Sakata, S. Lopez-Galarza, J. V. Maroto, S. G.
Lee, Y. C. Huh, Z. Sun, A. Miguel, S. R. King, R. Cohen, and J. M. Lee. 2008b.
Cucurbit grafting. Crit. Rev. Plant Sci. 27:50–74.
20. Davis, A. R., P. Perkins-Veazie, R. Hassell, S. R. King, and X. Zhang. 2008. Grafting
effects on vegetable quality. HortScience 43:1670–72.
21. Dillon, D. 1967. Simultaneous grafting and rooting of citrus under mist. Comb. Proc.
Intl. Plant Prop. Soc. 17:114–17.
22. Douglas, G. C., and J. McNamara. 1996. A tube method for grafting small diameter
scions of the hardwoods Quercus, Fraxinus, Betula and Sorbus. Comb. Proc. Intl.
Plant Prop. Soc. 46:221–26.
23. Eliezer E. Goldschmidt.2014. Plant grafting: new mechanisms, evolutionary
implications . TheRobert H. Smith Institute of Plant Sciences and Genetics in
Agriculture, Faculty of Agriculture, Food and Environment, The Hebrew University
of Jerusalem, Rehovot, Israel . Doi: 10.3389/fpls.2014.00727
24. Eichelser, J. 1967. Simultaneous grafting and rooting techniques as applied to
Rhododendrons. Comb. Proc. Intl. Plant Prop. Soc. 17:112.

29
25. Fisher, E. 1977. The pre-budded interstem: A new technique. Fruit Var. J. 31:14–5.
26. Flemer, W., III. 1986. New advances in bench grafting. Comb. Proc. Intl. Plant Prop.
Soc. 36: 545–49. 18. Gaggini, J. B. 1985. Bench grafting of trees under polyethylene.
Comb. Proc. Intl. Plant Prop. Soc. 34:646–47.
27. Garner, R. J. 1988. The grafter’s handbook. 5th ed. New York: Oxford Univ. Press.
28. Garner, R. (1958). Grafter's Handbook. New York: Oxford University Press. pp. 79–
100.
29. Garner, R. (1988). Grafter's Handbook. Cassell Illustrated. p. 131. ISBN 978-0-304-
32172-8.
30. Gisbert-Mullor R., Ceccanti C., Padilla Y.G., López-Galarza S., Calatayud A., Conte
G., and Guid L. 2020.Effect of Grafting on the Production, Physico-Chemical
Characteristics and Nutritional Quality of Fruit from Pepper Landraces. doi:
10.3390/antiox9060501.
31. Halma, F. F., and E. R. Eggers. 1936. Propagating citrus by twig-grafting. Proc.
Amer. Soc. Hort. Sci. 34:289–90.
32. Honma, S. 1977. Grafting eggplants. Scientia Hort. 7:207–11.
33. Hassell, R. L., F. Memmott, and D. G. Liere. 2008. Grafting methods for watermelon
production. HortScience 43:1677–9.
34. Hartmann, H.T.; Kester, D.; Davies, F.; Geneve, R. (2001). Plant Propagation:
Principals and Practices (https://archive.org/details/plantpropagation00hart) (7th ed.).
Prentice Hall. ISBN 978-0- 136-79235-2.
35. Harada T. 2010. Grafting and RNA transport via phloem tissue in horticultural plants.
Scientia Horticulturae 125: 545–550.
36. Hearman, J., A. B. Beakbane, R. G. Hatton, and W. A. Roach. 1936. The
reinvigoration of apple trees by the inarching of vigorous rootstocks. J. Pom. Hort.
Sci. 14:376–90.
37. Hottes, A. C. 1937. Plant propagation: 999 questions answered. New York: De La
Mare Co.
38. Hottes, A.C. (1925). Practical plant propagation: an exposition of the art and science
of increasing plants as practiced by the nurseryman, florist and gardener. New York:
A.T. De La Mare.

30
39. Howard, G. S., and A. C. Hildreth. 1963. Induction of callus tissue on apple grafts
prior to field planting and its growth effects. Proc. Amer. Soc. Hort. Sci. 82:11–5.
40. Huang, S., and D. F. Millikan. 1980. In vitro micrografting of apple shoot tips.
HortScience 15:741–43.
41. Intven, W. J., and T. J. Intven. 1989. Apical grafting of Acer palmatum and other
deciduous plants. Comb. Proc. Intl. Plant Prop. Soc. 39:409–12.
42. Jonard, R., J. Hugard, J. Macheix, J. Martinez, L. Mosella-Chancel, J. Luc Poessel,
and P. Villemur. 1983. In vitro micrografting and its application to fruit science.
Scientia Hort. 20:147–59.
43. Jackson S.D. (2009) Plant responses to photoperiod. New Phytol. 181: 517–531
44. Kerr, W. L. 1935. A simple method of obtaining fruit trees on their own roots. Proc.
Amer. Soc. Hort. Sci. 33:355–57.
45. Kubota, C., M. A. McClure, N. KokalisBurelle, M. G. Bausher, and E. N. Rosskopf.
2008. Vegetable grafting: History, use, and current technology status in North
America. HortScience 43:1664–9.
46. Kurata, K. 1994. Cultivation of grafted vegetables II. Development of grafting robots
in Japan. HortScience 29:240–44.
47. Kumar,G.(2011)."Propagation of Plants by Grafting and Budding"
(http://extension.oregonstate.edu/deschutes/sites/default/files/propagation.pdf) (PDF).
Pacific Northwest Extension. pp. 3–5.
48. Kollmann R, Glockmann C. 1985. Studies on graft unions. I. Plasmodesmata between
cells of plants belonging to different unrelated taxa. Protoplasma 124: 224–235
49. Kubota C., McClure M.A., Kokalis-Burelle N., Bausher M.G., Rosskopf E.N. (2008)
Vegetable graing: history, use, and current technology status in North America.
HortScience 43: 1664–1669.
50. Lagerstedt, H. B. 1982. A device for hot callusing graft unions of fruit and nut trees.
Comb. Proc. Intl. Plant Prop. Soc. 31:151–59.
51. Lagerstedt, H. B. 1984. Hot callusing pipe speeds up grafting. Amer. Nurs. 160:113–
17. 33. Lee, J. M. 1994. Cultivation of grafted vegetables 1. Current status, grafting
methods, and benefits. HortScience 29:235–39.

31
52. Li Yan , Sun Wei , Liu Fulin , Cheng J., Zhang Xiaojie , Zhang H., and Shandong
Y.Z .2019.Methods for grafting Arabidopsis thaliana and Eutrema salsugineum .
Provincial Key Laboratory of Plant Stress Research, College of Life Science,
Shandong Normal University, Jinan, 250014 Shandong China.doi: 10.1186/s13007-
019-0477-x .
53. Lee, J. M., and M. Oda. 2003. Grafting of herbaceous vegetable and ornamental
crops. Hort. Rev. 28:61–124.
54. Legare, M. 2007. The future of grafting. Comb. Prod. Intl. Plant Proc. Soc. 57:380–
84.
55. Leiss, J. 1987. Modified side graft for nursery trees. Comb. Proc. Intl. Plant Prop.
Soc. 36:543–44.
56. Lincoln, F. B. 1938. Layering of root grafts—a ready method for obtaining self-
rooted apple trees. Proc. Amer. Soc. Hort. Sci. 35:419–22.
57. Langford, M. H., and J. C. H. T. Townsend, Jr.. 1954. Control of South American leaf
blight of Hevea rubber trees. Plant Dis. Rpt. Suppl. p. 225.
58. Lee, J. M.1994. Cultivation of grafted vegetables. I. Current status, grafting methods,
and benefits. HortScience 29:235–39.
59. Lee, J. M., and M. Oda. 2003. Grafting of herbaceous vegetable and ornamental
crops. Hort. Rev. 28:61–124.
60. Loehle, C. & Jones, R.H. (1998). "Adaptive Significance of Root Grafting in Trees".
Functional Ecology. 4 (2): 268–271. JSTOR 2389347.
61. MacDonald, B. 1986. Practical woody plant propagation for nursery growers.
Portland, OR: Timber Press.
62. Marcelo de A Guimarães, Jeffery L Gustin, and David E Salt 2009 . Reciprocal
grafting separates the roles of the root and shoot in zinc hyperaccumulation in Thlaspi
caerulescens.Department of Horticulture and Landscape Architecture, Purdue
University, West Lafayette, IN 47906, USA.. doi: 10.1111/j.1469-8137.2009.02969.x
63. McAdam S.A.M., Brodribb T.J., Ross J.J. (2016) Shoot-derived abscisic acid
promotes root growth. Plant Cell Environ. 39: 652–659
64. Melnyk CW, Meyerowitz EM. 2015. Plant grafting. Current Biology 25: R183– R188

32
65. McEachern, G. R., and A. Stockton. 1993. The four-flap graft. In G. R. McEachern
and L. A. Stein, eds. Texas pecan handbook. TAEX Hort Handbook 105. College
Station, TX: Texas Agricultural Extension Service, Texas A&M University.
66. McEachern, G. R., S. Helmers, L. Stein, J. Lipe, and L. Shreve. 1993. Texas inlay
bark graft. In G. R. McEachern and L. A. Stein, eds. Texas pecan handbook. TAEX
Hort Handbook 105. College Station, TX: Texas Agricultural Extension Service,
Texas A&M University.
67. Maynard, B. K., and N. L. Bassuk. 1990. Comparisons of stock plant etiolation with
traditional propagation methods. Comb. Proc. Intl. Plant Prop. Soc. 40:517–23.
68. McClintock, J. A. 1948. A study of uncongeniality between peaches as scions and the
Marianna plum as a stock. J. Agr. Res. 77:253–60.
69. Paultre DSG, Gustin M-P, Molnar A, Oparka KJ. 2016. Lost in transit: longdistance
trafficking and phloem unloading of protein signals in Arabidopsis homografts. The
Plant Cell 28: 2016–2025
70. Raulston, J. C. 1995. New concepts in improving ornamental plant adaptability with
stress tolerant rootstocks. Comb. Proc. Intl. Plant Prop. Soc. 45:566–69.
71. Richards, D., W. K. Thompson, and R. P. Pharis. 1986. The influence of dwarfing
interstocks on the distribution and metabolism of xylem-applied [3 H] Gibberellin A4
in apple. Plant Physiol. 82:1090–95. 131. Rivard, C. L., and F. J. Louws. 2008.
Grafting to manage soilborne diseases in heirloom tomato production. HortScience
43:2104–11.
72. Sharma A. and Zheng B..2019. Molecular Responses during Plant Grafting and Its
Regulation by Auxins, Cytokinins, and Gibberellins. State Key Laboratory of
Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, China.
doi:10.3390/biom9090397
73. S. J. Cookson, M. J. Clemente Moreno, C. Hevin, L. Z. Nyamba Mendome, S. Delrot,
N. Magnin, C. Trossat-Magnin, and N. Ollat 2014 . Heterografting with nonself
rootstocks induces genes involved in stress responses at the graft interface when
compared with autografted controls . doi: 10.1093/jxb/eru145.

33
74. Slagman S., Prof. Dr. Zuilhof H., and Dr. Maurice C. R. Franssen 2017.
Laccase‐Mediated Grafting on Biopolymers and Synthetic Polymers: A Critical
Review. doi: 10.1002/cbic.201700518
75. Stratmann J.W. (2003) Long distance run in the wound response—jasmonic acid is
pulling ahead. Trends Plant Sci. 8: 247–250
76. "Terms and Conditions of Supply of Budwood by CCPP"
(http://ccpp.ucr.edu/budwood/budwood.php). Citrus Clonal Protection Program.
University of California, Riverside. Retrieved 13 September 2017.
77. Tsutsui H. and Notaguchi M..2017.The Use of Grafting to Study Systemic Signaling
in Plants .Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho,
Chikusa-ku, Nagoya, Aichi 464-8601, Japan 2 Japan Science and Technology
Agency, PRESTO, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan
.doi:10.1093/pcp/pcx098
78. Wang J., Jiang L. and Rongling Wu.2016. Plant grafting: how genetic exchange
promotes vascular reconnection. Center for Computational Biology, College of
Biological Sciences and Technology, Beijing Forestry University, Beijing 100083,
China; 2 Center for Statistical Genetics, Pennsylvania State University, Hershey, PA
17033, USA .doi: 10.1111/nph.14383
79. Warschefsky EJ, Klein LL, Frank MH, Chitwood DH, Londo JP, von Wettberg EJ,
Miller AJ. 2015. Rootstocks: diversity, domestication, and impacts on shoot
phenotypes. Trends in Plant Science 21: 418–437
80. Willingham S, Pegg K, Cooke A, Coates L, Langdon P, Dean J. 2001. Rootstock
influences postharvest anthracnose development in ‘Hass’ avocado. Crop and Pasture
Science 52: 1017–1022

34