The research is based on epicotyl grafting of mango

2. INFLUENCE OF ROOTSTOCK AGE AND
PRE-DEFOLIATION OF SCION ON THE
SUCCESS OF EPICOTYL GRAFTING OF
MANGO
Principal Investigator:
Bijaya Upadhya
R-2011-HRT-07M
Department of Horticulture

3. Advisory Committee
Chairman: Prof. Dilli Ram Baral, Ph.D.
Department of Horticulture
IAAS, Rampur, Chitwan
Members: 1. Prof. Durga Mani Gautam, Ph.D.
Department of Horticulture
IAAS, Rampur, Chitwan
2. Prof. Sundarman Shrestha, Ph. D
Department of Pathology
IAAS, Rampur, Chitwan

4. •Mango (Mangifera indica) is one of the leading fruit crops of Nepal.
•Covers 9% of total cultivated area of horticultural crops
•Area – 40, 110 ha
•Production – 3, 28, 883 mt
• Productivity – 10.21 mt/ha (MOAC, 2011/12)
•Productivity is lower than that in India and other countries
•Based on this scenario and opportunities, five years plan have
emphasized on commercial orcharding of mango in 15 districts.
•Nature of crop – cross pollination
•Mango is commercially propagated vegetatively due to its heterozygous
nature.
Introduction

5. Cont….
•Grafting is the commercial method for mango propagation in Nepal.
•Epicotyl grafting or Stone grafting of mango was first reported by Traub
and Auchter (1934).
•Success in epicotyl grafting may vary depending upon age of rootstock,
variety of scion, pre-defoliation of scion, length of scion, grafting
season etc.
•Lack of Healthy planting
material.
•Lack of improved technology in
farmers level.
•Lack of reliable and reasonable
planting material.
•Propagation through sexual
method are not true to type.
•Prevailing grafting technology
inarching is difficult, time
consuming and laborious.
• Epicotyl grafting: higher success
rates, are more cost efficient,
because of lower maintenance
costs, and result in more rapid
multiplication of vigorous and
healthy grafted seedlings.
•Pre-defoliation of scion cause an
immediate rise in sucrose content
of phloem sap of the shoots.
•Rootstock contain higher
amount of sap and meristematic
activity is high.
Problems Rationale of study

6. General objective
To evaluate the effect of age of root stock and pre-defoliation
of scion on success of epicotyl grafting in mango.
Specific objective
•To determine the effect of age of rootstock on success of
epicotyl grafting in mango
•To find out the effect of pre-defoliation of scion on success of
epicotyl grafting in mango
•To find out the interaction effect of age of rootstock and pre-
defoliation of scion on success of epicotyl grafting in mango
Objectives

7. •Origin
• Botanical description
• Importance and global scenario of mango
• Grafting technology/graftage
• Epicotyl grafting
• Pre-defoliation of scion
• Age of rootstock in epicotyls grafting of mango Climatic factor affecting
success of Epicotyl grafting
•Season of grafting
• Survival rate and mortality
• Anatomy of graft union
Literature review

9. Materials and Methods
Location and Site of experiment
Research site and techniques used
Study area and its location Shade house of horticulture department of
IAAS
Duration of experiment June – November 2012
Grafting techniques Epicotyl grafting
Method of Epicotyl grafting Cleft grafting
Rampur, Chitwan,
Nepal

10. 0
100
200
300
400
500
600
0
10
20
30
40
50
60
70
80
90
100
June July August September October November
Rainfall / mm Maximum Minimum Relative humidity (%)
Figure 1. Agro-meterological data recorded during June-November, 2012 at Rampur,
Chitwan, Nepal

11. Preparation of rootstock:
•Fruits from 10-15 years old local mango variety was collected
for preparation of rootstock. The mean weight of ripe fruit
taken was 56gm.
• Seed stones were extracted from fruits and kept in water for
24hrs for fermentation and washed in tap water properly and
placed under shade.
•200 Seed stones were sown each in 24th, 29th of June and 4th
and 9th of July. Altogether 800 seed stones were sown.
Germination of seed was taken on 7-8 days
•Media preparation: soil : saw dust:: 1:1 Mango seed stone
5 days old seedling 10 days old seedling 15 days old seedling 20 days old seedling

12. Saw dust Soil
Diameter of rootstock taken Length of rootstock taken

13. Preparation of scion
 3-4 month old pre-defoliated healthy scion of Dasehari mango
were collected.
Scion was defoliated in 7th, 12th, 17th and 22nd of July 2012.
Pre-defoliation was done by removing off the leaf with sharp
scissor and retaining leaf petiole on the mother tree in the
morning time of the day
Fig: Pre-defoliation of scion
Length of scion taken

14. •Grafted plants were covered by
polythene cap to maintain high
humidity
•0.2% solution of bavistin was
sprayed at regular intervals of 4
days on the grafted plants.
•Suckers were removed as per
requirement
•Experimental setup: Two factorial
RCBD
•Replication: 3
Grafted plant covered with polythene cap

15. Treatment Detail: 16 treatments were incorporated
Factor A: ( Age of rootstock)
A1- 5 days old seedling
A2- 10 days old seedling
A3- 15 days old seedlings
A4-20 days old seedlings
Factor B: (Pre-defoliation of scion)
D1- 0 Days before grafting
D2- 5 Days before grafting
D3- 10 Days before grafting
D4- 15 days before grafting
Each treatment consists of 10 plants in every replication

16. Layout of Research
R1
R2
R3

17. Observation taken:
 Days to first sprouting
 Days to complete sprouting
 Initial success
 Final Success
 Mortality of graft
 Shoot length
 Shoot diameter
 No of growth flushes
 Leaf length
 Leaf width
 Height of graft
 Spread of graft
 Volume of graft

18. Data analysis and interpretation
Data entry was done in Microsoft excel and Statistical analysis was
done by using statistical software program MSTAT-C and Microsoft
excel. Means were further separated by DMRT where treatment
results were found significant.

19. Result and Discussion

20. Table 1. Effect of rootstock age and pre-defoliation of scion on sprouting and success
of the grafts in epicotyl grafting of mango at IAAS, Rampur, 2012
Treatments
Days to first
sprouting
Days to completion of
sprouting
Initial success (%)
Final success
(%)
Factor A (Rootstock age)
5 Days 13.58 a 19.58 a 23.33d 12.50 d
10 Days 13.17a 19.33 ab 46.67 c 24.17 c
15 Days 12.5 b 18.83b 65.00 a 38.33a
20 Days 13.67a 20.00 a 56.67 b 33.33 b
LSD 0.634** 0.704* 0.527** 0.461**
SEm (±) 0.219 0.243 0.182 0.159
Factor B (Pre-defoliation of scion)
0 Days 15.42a 21.50 a 39.17 c 20.00 c
5 Days 14.00 b 20.17 b 45.83 b 26.67 b
10 Days 11.17d 17.42d 56.67 a 39.17 a
15 Days 12.33 c 18.67c 50.00 b 22.50 bc
LSD 0.634** 0.704** 0.526** 0.461**
SEm 0.219 0.243 0.182 0.159
CV (%) 5.75 4.34 13.18 20.41

21. Treatments
Days to first
sprouting
Days to completion of
sprouting
Initial success (%) Final success (%)
A1D1 16.00a 22.00a
16.7i 6.70h
A1D2 14.67a 20.67ab
26.7hi 13.3fgh
A1 D3 11.00c 16.67d
20.0hi 20.0efg
A1D4 12.67b 19.00c
30.0gh 10.0gh
A2D1 15.00a 21.00a
40.0fg 13.3fgh
A2D2 13.33b 19.33bc
43.3ef 23.3def
A2D3 12.00bc 18.33c
53.3cde 33.3cd
A2D4 12.33bc 18.67c
50.0def 26.7cde
A3D1 15.00a 21.33a
50.0def 36.7c
A3D2 13.00b 19.00c
60.0bcd 33.3cd
A3D3 9.667d 16.33d
86.7a 56.7a
A3D4 12.33bc 18.67c
63.3bc 26.7cde
A4D1 15.67a 21.67a
50.0def 23.3def
A4D2 15.00a 21.67a
53.3cde 36.7c
A4D3 12.00bc 18.33c
66.7b 46.7b
A4D4 12.00bc 18.33c
56.7bcd 26.7cde
LSD 1.269* 1.408* 1.053** 0.922*
SEm (±) 0.439 0.487 0.365 0.319
CV (%) 5.75 4.34 13.18 20.41
Table 2. Interaction effect of rootstock age and pre-defoliation of scion on sprouting and
success of the grafts in epicotyl grafting of mango at IAAS, Rampur, 2012

22. y = 1.0908x + 18.373
R² = 0.6608
0
10
20
30
40
50
60
70
80
90
100
0 10 20 30 40 50 60 70
Initial
success
(%)
Final success (%)
Figure 16. Correlation between initial and final success percentage of graft in epicotyl
grafting of mango

23. •Reddy and Kohli (1989) recorded maximum success (66.66%)
from 14 days old rootstocks in Alphonso. Also, Alam et al.
(2006) revealed that maximum final success was obtained on
15 day old seedlings followed by 20 day old seedlings.
•Dhungana et al. (1989) reported that highest percentage of
survival (49.50%) was observed when scions were defoliated 10
days prior to epicotyl grafting compared to 15 days prior to
defoliation (47.50%) or 0 days pre-defoliation (33.50%) under
Kerala condition.

24. 10.8
22.5 26.7 23.3
76.7 53.3
35 43.33
0
10
20
30
40
50
60
70
80
90
100
5 Days 10 Days 15 Days 20 Days
45 DAG 90 DAG
Rootstock age
Mortality
of
grafts
(%)
Figure 1. Effect of rootstock age on mortality rate (%) of grafts at 45 and 90 days after epicotyl
grafting in mango at IAAS, Rampur, 2012

25. 19.2 19.2 17.5
27.5
60.8
54.1
43.3
50
0
10
20
30
40
50
60
70
80
90
0 Days 5 Days 10 Days 15 Days
45 DAG 90 DAG
Mortality
of
grafts
(%)
Pre-defoliation of scion
Figure 2. Effect of pre-defoliation of scion on mortality rate (%) of grafts at 45 and 90 days
after epicotyl grafting in mango at IAAS, Rampur, 2012

26. 0
10
20
30
40
50
60
70
80
90
A1D1 A1D2 A1 D3 A1D4 A2D1 A2D2 A2D3 A2D4 A3D1 A3D2 A3D3 A3D4 A4D1 A4D2 A4D3 A4D4
Mortality
(%)
Treatment combination
Figure 3. Interaction effect of rootstock age and pre-defoliation of scion on mortality
rate (%) of grafts at 45 days after grafting on epicotyl grafting of mango at IAAS,
Rampur, 2012

27. Treatments
Shoot
length(cm)
Shoot
Diameter
(cm)
Number of
Growth
flushes
Number of
leaves
Leaf length
(cm)
Leaf width
(cm)
Spread of
graft (cm)
Factor A
A1 17.84 c 0.50 b 1.41 c 8.41 b 13.73 b 2.10 b 24.46 c
A2 21.25 b 0.56 ab 2.16 b 12.17 a 16.42 a 2.27 a 25.88 ab
A3 22.34 a 0.58 a 2.75 a 13.25 a 17.39 a 2.30 a 26.61 a
A4 20.53 b 0.55 ab 2.08 b 12.50 a 15.52 ab 2.15 ab 25.61 b
LSD 0.721** 0.052* 0.475** 1.125** 1.979** 0.158* 0.902**
SEm(±) 0.249 0.178 0.164 0.389 0.685 0.055 0.312
Factor B
D1 19.58 c 0.49 c 1.50c 10.08 b 14.21 b 2.02 b 24.54 c
D2 20.51 b 0.56ab 2.25 b 12.25 a 16.04 ab 2.19 a 25.92 b
D3 21.88 a 0.60 a 2.75 a 12.33 a 17.08 a 2.34 a 27.06 a
D4 19.99 bc 0.54 bc 1.91 bc 11.67 a 15.74 ab 2.29a 25.02 bc
LSD 0.721** 0.052** 0.475** 1.125** 1.979* 0.158** 0.902**
SEm (±) 0.249 0.178 0.164 0.389 0.685 0.055 0.312
CV (%) 4.22 11.16 27.04 11.65 15.05 8.62 4.22
Table 3. Effects of rootstock age and pre-defoliation of scion on growth parameters of the
grafts in epicotyl grafting of mango at IAAS, Rampur, 2012

28. Treatments SL (cm) SD (cm) NOGF NOL LL(cm) LW(cm) SOG (cm)
A1D1 17.07h 0.46 1.00 7.00h 12.90 1.90 22.60f
A1D2 16.93h 0.50 1.33 8.00gh 14.00 2.03 26.23bcd
A1 D3 19.27efg 0.55 2.00 10.00efg 14.40 2.27 25.37bcde
A1D4 18.10gh 0.51 1.33 8.67fgh 13.60 2.23 23.63ef
A2D1 18.77g 0.49 1.67 10.00efg 14.77 2.03 23.83ef
A2D2 22.07bcd 0.59 2.33 13.67abc 16.80 2.37 26.73abcd
A2D3 22.80ab 0.61 2.67 11.00def 17.73 2.43 27.40ab
A2D4 21.37bcd 0.54 2.00 14.00ab 16.40 2.27 25.53bcde
A3D1 21.97bcd 0.53 1.67 11.33cde 15.43 2.20 26.83abcd
A3D2 22.27abc 0.60 3.00 13.67abc 17.73 2.13 25.57bcde
A3D3 23.77a 0.65 3.67 15.33a 18.97 2.40 28.33a
A3D4 21.37bcd 0.56 2.67 12.67bcd 17.43 2.50 25.70bcde
A4D1 20.53def 0.51 1.67 12.00bcde 13.73 1.93 24.90de
A4D2 20.77cde 0.55 2.33 13.67abc 15.63 2.23 25.17cde
A4D3 21.70bcd 0.60 2.67 13.00abcd 17.20 2.27 27.13abc
A4D4 19.13fg 0.55 1.67 11.33cde 15.53 2.17 25.23cde
LSD 1.42** NS NS 2.25* NS NS 1.80*
SEm (±) 0.499 - - 0.779 - - 0.625
CV% 4.22 11.16 27.04 11.65 15.05 8.62 4.22
Table 4. Interaction effects of rootstock age and pre defoliation of scion on growth parameters
of the grafts in epicotyl grafting of mango at IAAS, Rampur, 2012

29. • Alam et al. (2006) reported that all the growth parameter was
observed maximum in 15 days old seedling which was similar to
current study.
• The maximum growth in the grafts of 10 days pre-defoliated scion
might be due to optimum amount of sucrose content in bud level
thereby initiation of higher meristamatic activity which helps in
better sap flow and good callus formation due to stimulation of
cambium division favoring better graft union (Maiti and Biswas,
1980).

30. Figure 4. Effect of rootstock age on graft height in epicotyl grafting of mango at
IAAS, Rampur, 2012
24.25
25.83
27.75
25.67
22
23
24
25
26
27
28
29
5 Days 10 Days 15 Days 20 Days
Graft
height
(cm)
Rootstock age

31. Figure 5. Effect of Pre-defoliation of scion on graft height in epicotyl grafting of
mango at IAAS, Rampur, 2012
24.5
26.17
27.08
25.75
23
23.5
24
24.5
25
25.5
26
26.5
27
27.5
0 Days 5 Days 10 Days 15 Days
Graft
height
(cm)
Pre-defoliation of scion

32. Treatments
Rootstock age
Pre-defoliation of Scion
0 Days 5 Days 10 Days 15 Days
5 Days 23.33 24.00 25.00 24.67
10 Days 24.67 26.00 26.67 26.00
15 Days 25.67 28.33 30.00 27.00
20 Days 24.33 26.33 26.67 25.33
DMRT NS
CV (%) 6.73
Table 5. Interaction effect of rootstock age and pre-defoliated scion on graft height in
epicotyl grafting of mango at IAAS, Rampur, 2012

33. • The highest graft height with 15 days old seedling might be due
to earliest sprouting and better stock and scion relationship (Patil
et al., 1984).
• The maximum height of the graft (21.96cm) was found in 9 days
pre-defoliated scion while the minimum was obtained in un-
defoliated scion in guava plants (Kumar et al., 2012).

34. Figure 6. Effect of rootstock age on volume of graft in epicotyl grafting of mango at
IAAS, Rampur, 2012
3649
4354
4948
4227
2000
2500
3000
3500
4000
4500
5000
5500
5 Days 10 Days 15 Days 20 Days
Volume
of
graft
(cm
3
)
Rootstock age

35. Figure 7. Effect of pre-defoliation of scion on volume of graft in epicotyl grafting of
mango at IAAS, Rampur, 2012.
3727
4392
5004
4056
2000
2500
3000
3500
4000
4500
5000
5500
0 Days 5 Days 10 Days 15 Days
Volume
of
graft
(cm
3
)
Pre-defoliation of scion

36. Table 6. Interaction effect of rootstock age and pre-defoliation of scion on volume
of graft (cm3) in epicotyl grafting of mango at IAAS, Rampur, 2012
Treatments Pre-defoliation of scion
Rootstock age 0 Days 5 Days 10 Days 15 Days
5 Days 2987.06 4111.37 4055.23 3442.92
10 Days 3510.03 4654.12 5018.57 4235.24
15 Days 4627.74 4638.88 6028.82 4497.61
20 Days 3781.28 4163.54 4913.08 4048.62
DMRT NS
6.7
CV (%)

37. •Alam et al. (2006) reported that the maximum leaf area was
obtained in the graft of 15 days old seedlings.
•The maximum volume of graft might be due to high meristamatic
activity in the bud level which leads to maximum shoot length,
number of leaves, number of growth flushes, leaf length, graft
height and spread of graft resulting maximum volume of graft.

38. The maximum initial and final success of the grafted plant was
obtained in 15 days old seedlings and 10 days pre-defoliation of
scion where as minimum was found in the grafted plant of 5 days
old seedlings and un-defoliated scion.
The maximum growth parameter of the grafted plant was
obtained in 15 days old seedlings and 10 days pre-defoliation of
scion where as minimum was found in the grafted plant of 5 days
old seedlings and un-defoliated scion. Thus, 15 days old seedling
and 10 days pre-defoliation of scion does best in Chitwan
condition.
Summary and Conclusion

39. Research glimpse
Collection of seed stones
Seed stones kept in shade Pre-defoliation of scion

40. Germination media Media filling in polybags
Seed stones in Bucket Preparation of media in Polybags for
sowing

41. Seedlings of mango Grafted plant of mango
Grafted plant covered with
polythene cap
Humiditiy maintanance in grafted
plant

42. Succeed grafted plant
Sprouting in mango Sprouted graft

43. Advisor visit

44. Dean IAAS
Assistant deans
PG-program coordinator
Members of the advisory committee
Friends and colleagues
Family
Acknowledgement

45. Queries and questions are heartily welcome