Proceedings of the 4th International Symposium on Machinery and Mechatronics for

Agriculture and Biosystems Engineering (ISMAB)

27-29 May 2008, Taichung, Taiwan

Grafting Efficiency Evaluation of Sweet Pepper Grafted by a

Tubing-Grafting Robotic System
Yung-Chiung Chang1, Suming Chen2, Lian-Hsiung Lin3, and Yi-Chich Chiu4*
1

Dept. of Horticulture, National ILan University, Taiwan.

Dept. of Bio-Industrial Mechatronics Engineering, National Taiwan University, Taiwan.
3,4
Dept. of Biomechatronic Engineering, National ILan University, Taiwan.
*Corresponding Author, Email: yichiu@niu.edu.tw

Abstract: Seedlings growth conditions of grafted by the machine were as well as those by traditional
hand grafting. Although the cut interfaces of the robotic grafting were more irregular, the acclimation
process was similar to that of the hand operation. It indicates that the tubing-grafting robotic system
can be applied to grafting operations and improve the production efficiency.
Key Words: sweet pepper, grafting, robot, tubing, histology

INTRODUCTION
Grafting technique has been significantly improved during the last year. Vegetable grafting is
extremely popular in some Asiatic country (Lee, 1994), and in Europe the use of vegetable grafting is
now expanding, too (Fernndea-Garca et al., 2004). Because of having the advantages of reducing
infections caused by pathogens (Bils et al., 1989; Padgett and Morrison, 1990), increasing the
resistance to drought (White and Castillo, 1989) and enhancing nutrient uptake (Ruiz-Sifre et al.,
1997), the grafting technique is extensively applied in vegetable production. As high labor cost is still
the major problem of this property, attempts to mechanize grafting operation have been the important
subject.
Japan has actively been engaged in the development of grafting techniques. Grafting robotic
systems developed by the Biology Research Organization (Kobayashi and Suzuki, 1996; Kobayashi et
al. 1966) can do 1,200 grafts per hour and obtain the success rate of 98% for grafting. Nishiura et al.
(1995) developed a system of grafting robotic systems using the plug-in method. The system can
reduce the mismatch of vascular bundles during grafting, and thus can fasten the healing of grafted
seedlings and make seedlings more vigorous. In Korea, Hwang et al. (1997) developed a grafting
robotics for fruit-bearing vegetables based on inarching grafting. In Taiwan, Lee et al. (2001)
developed automatic graft robotic systems for grafting Passion Melon. It can graft 114 seedlings per
hour, and has the grafting success rate of 70% and survival rate of 95%. As for others like Kurata
(1994), Lee (1994), they have also conducted research on grafting robotic systems. As for the
commercialization of grafting robotic systems, three companies including YANMAR, ISEKI and
MITSUBISHI, assisted by the Biology Research Organization, have developed a grafting robotic
system respectively in Japan, which can do automatic grafting for cucurbitaceae and Solanaceae plants,
with operation capability of 550~1000 seedlings
per hour and a success rate of 95%.
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Some methods are used to confirm the functions of the grafting robots. The grafted plants
vigorous and the grafted unions acclimating process are the most obvious appearances to differentiate
the success or failure of the grafting operations. Hartmann et al. (2002) described the sequence events
of the grafted herbaceous plants. First, new parenchymatous cells proliferate from both stock and scion
produce the callus tissues that fill up the spaces between the two components connecting the scion and
the stock. Following, the new cambial cells differentiate from the newly formed callus, forming a
continuous cambial connection between the stock 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. Finally, 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.
As the necessity of improving the grafting operation efficiency, in this study, we investigated the
performances of the tubing-grafting robotic system and went a step further to discuss the vigorous and
acclimating process of the grafted plants. We chose the sweet peppers for the examined plants,
because they not only an important vegetable used widely but also their grafted seedlings have been
utilized more and more extensively in the worldwide.

MATERIALS AND METHODS

Design of the tubing-grafting robotic system
The tubing-grafting robotic system is composed of six major units: the chucking and fetching
rootstocks, the cutting rootstocks, the chucking and fetching scions, the cutting scions, the supplying
and cutting rubber tubes, and the guiding tubing. It is programmatically controlled by programmable
logic controllers cooperated with pneumatic driving mechanisms. The design and structure of the
tubing-grafting robotic system is as shown as the Fig.1.

Feeding wheel of tube

Scion cutting unit

Chucking and fetching
Scion unit

Tube supplying and

cutting unit

Rootstock cutting unit

Guiding grafting unit

Chucking and fetching
rootstock unit

Pneumatic source

Control panel
Figure 1. The design of the tubing-grafting robotic system

Plant material
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In this research, the sweet pepper Andalus (Capsicum Annuum L. Andalus) were used as scions
and the chili pepper P (Capsicum Chinense P)were used as stocks. Seedlings used in the experiment
were all purchased from Yui-Cha Seedling Nursery where was located at Chiayi county. The average
sizes of the tested seedlings were 14 cm height and borne 3-4 unfolded true leaves.
On 3 Aug. 2007, sweet peppers were grafted on chili peppers with the tubes of 2 mm diameters by
robotic method or hand. And both manually and mechanically grafting methods had 128 plants grafted,
respectively. After grafted, the grafted plants were transferred to an acclimatization chamber maintained
at 27.51.5 and 905 % relative humidity (RH) under 49.12 mol/m2/s photosynthetic photon flux
light intensity (lights turned on from 8 AM to 8 PM during experimental period). Four days later, grafted
seedlings were moved from the chamber into the tunnel shelter for further acclimation.
After acclimation, the grafted plants were transferred to the greenhouse. On 17 Aug. all grafted
plants were transplanted into plastic pots which were 1000 mL and filled with median of BVB. A
slow-released granular fertilizer with N, P, and K each at 20, 20, and 20 g/m2 had been incorporated
into the median before transplanted. Besides, normal cultural practices for each experiment were
followed for irrigation, fertilizer, and pesticide application.
In this part, a completely randomized design (CRD) was used. The survival ratio was recorded
each two days from the 1st day after grafted (DAG) to the 13th. Plants growing conditions were
recorded too, including the variance of height and diameter for the stocks and scions. All data
collected from the experiments were subjected to analysis of variance (one way ANOVA) using SAS
package software (SAS Institute, 1996) and LSD (p = 0.05) calculated.

Union acclimation process of grafted plants

The samples of both robot and hand grafted combinations were harvested every day after grafting
0-14 days, and each two days during grafting 15-56 days, respectively. 6 samples were harvested at
every detecting day, and were used for plant histology observation by paraffin sections. The specimens
were trimmed to 5 mm above and below the graft union (1.0 cm long), fixed for 24 h in
formalin-acetic-ethanol (FAA), dehydrated through a series of graded ethanol-tertiary-butyl alcohol
(TBA) series and two times in absolute ethanol for 2 h per stage. Tissue was then embedded in paraffin
in an oven at 55. The fixed samples were sectioned longitudinally with a rotator microtome (Rotary
microtome, N. O. W 212, Japan) at a thickness of 15 m, and then placed on glass slides. The sections
were then deparaffinized in xylene, rehydrated in an ethanol series for 2 min (100%, 70%, and 40%)
and washed in distilled water. Sections were stained with 1 % aqueous safranin O (Sigma) for 8 hours
and 0.5 % fast-green (Sigma) in 95 % ethanol for another 8 hours.

RESULTS AND DISCUSSION

Effect of grafting methods on growth of sweet pepper
The variance of seedlings survival ratio is as shown as the Table 1. After grafted by both hand
and robot, all sweet pepper seedlings wilted apparently in the previous days after grafting. After 5 days
of grafting, 80 % of the hand grafting seedlings recovered and two days later all seedlings were
vigorous. Although the recovering process of the robotic grafting seedlings was slower than that of the
hand, after 7 days of grafting, seedlings of robotic grafting were all visible vigor. The variances of
height in stock (chili pepper) were not obvious during the experimental period, even though there were
significant differences between the two grafting methods. The growth curves of scions were similar in
both grafting methods, as the robotic seedlings had significant higher height than those of hand at the
previous 14 days after grafting (Fig. 2).
Table 1. Effect of different grafting methods on survival ratio of sweet pepper grafted seedling
Grafting
Survival ratio (%)
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1 DAG z
0y
0

method
Hand
Robot

3 DAG
0
0

5 DAG
80
0

7 DAG
100
100

9 DAG
100
100

11 DAG
100
100

13 DAG
100
100

: DAG, days after grafting.

: Each data was the percentage of the recovered seedlings per 30 grafted seedlings.

22
20

robot
hand

Height of stock (mm)

18
16
14
12
10
8

14

21

28

35

42

49

56

28

35

42

49

6
4
2
0
22
20

robot
hand

Height of scion (mm)

18
16
14
12
10
8

6
4

2
0

2007 Agu. 3

14

21

Agu. 31

56
Sep. 28

Days after grafting

Figure 2. Effect of different grafting methods on the height of the sweet pepper grafted seedlings. A,
and B are height of stock and scion, respectively. Data are the mean of 30 plants. Vertical
bars represent LSD at 5%. The different letters by the symbols are significantly different at
the 5 % level (by LSD).

Acclimation process of the grafted union of sweet pepper

Fig. 3A to 3D illustrates the histology of union acclimation processes of the hand grafting. On 1
DAG, the graft interval between stock and scion was apparently but the cuts were smooth and regular
(Fig. 3A). On 5 DAG, some new proliferated callus tissue filled in the graft gap and formed a callus
bridge that made scion and stock connected (Fig. 3B). On 12 DAG, new vascular cambium cells
differentiated (Fig. 3C). On 20 DAG, the combining phenomena were clear and definite and appeared
by the restoration of new vascular tissue which went round to meet the target tissues therefore made a
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swelling portion around the cut (Fig. 3D).

The graft unions histology of the sweet pepper seedlings grafted by tubing-grafting robotic
system are as shown as Fig. 3E to 3H. On 1 DAG, the obvious graft interval was also observed
however the cuts were uneven and irregular which were not the same as the hand grafting ones (Fig.
3E). On 5 DAG, necrotic layer covered the interfaces and large new proliferated callus tissue filled in
the graft gap (Fig. 3F). On 12 DAG, the new vascular cambium cells formed though there were large
fractions around the interfaces (Fig. 3G). On 20 DAG, new vascular tissues restored and because of the
uneven cuts large callus proliferated and formed a swelling tumor around the cut (Fig. 3H).
For the majority of authors, the vascular connection of scion and stock is considered as the basic
requirement for a successful graft (Moore and Walker, 1981a, Moore and Walker, 1981b, Yeoman,
1984). Although the graft interfaces of mechanical grafting seedlings were more uneven and irregular
than those of hand grafting, the evidences of vascular connection between two plants were still definite.
As the growth performances of the mechanical grafted seedlings were similar with the hand grafting
ones, its no doubt that the tubing-grafting robot system can be utilized as a functional machine to
improve the grafting efficiency.

CONCLUSIONS
In this study, the tubing-grafting robotic system appeared a good function on sweet pepper
grafting. Growth condition of seedlings grafted by the machine was as well as those by traditional
hand grafting. Although the cut interfaces of the robotic grafting were more irregular, the acclimation
process was similar to that of the hand operation. It indicates that the tubing-grafting robotic system
can be applied to grafting operations and improve the production efficiency.

REFERENCES
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Fernndea-Garca, N., M. Carvajal , and E. Olmos. 2004. Graft union formation in tomato plants: peroxidase and
catalase involvement. Ann. Botany 93: 53-60.
Hartmann, H. T., D. E. Kester, F. T. Davies, and R. L. Geneve. 2002. Plant Propagation. Principle and Practices,
seventh ed. Prentice Hall, Upper Saddle River, NJ, 849 pp. ISBN 0-13-679235-9.
Hwang, H., J. H. Chang, and S. C. Kim. 1997. Automatic grafting system for fruit bearing vegetables based on
inarching graftage. Proceedings of ISAMA 97:75-80, Taipei: Taiwan.
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vegetables (part 4). J. Jap. Agr. Machinery 58(3): 59-68.
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Ruiz-Sifre, G., L. R. Santiago-Santos, and L. V. Ramirez-Ramos. 1997. Bioregulators and poinsettia plant
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453-472.

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Figure 3. Acclimation processes of the graft union of sweet pepper seedlings grafted by hand (A to
D) and robotic (E to H) grafting. (A) and (E) Longitudinal section of the graft union on 1
DAG, showing the apparent interval between stock and scion. (B) and (F) Longitudinal
section of the graft union on 5 DAG, new parenchymatous cells proliferate from both
stock and scion producing the callus tissue and intermingles and interlocks. (C) and (G)
Longitudinal section of the graft union on 12 DAG, new forming callus were filled up the
spaces between the two components connecting the scion and stock. (D) and (H)
Longitudinal section of the graft union on 20 DAG, new cambial cells differentiate from
the newly formed callus, forming a continuous cambial connection between scion and
stock.

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