EVALUATION OF SUITABILITY OF

DIFFERENT CUTTING TYPES AND TISSUE

CULTURE PLANTS, AS PLANTING

MATERIALS FOR VANILLA (Vanilla fragrans

L.) CULTIVATION.

C. I. M. Attanayake

Central Research Station

Department of Export Agriculture,

Matale

INTRODUCTION

Vanilla - tropical climbing orchid

Family - Orchidaceae

110 species were described in the world, Commercially

important species are

1. Vanilla fragrans

2. Vanilla pompona

3. Vanilla tahitensis

Most economically important and commercially

cultivated spice - Vanilla fragrans

….INTRODUCTION

Importance: Extract popular flavoring substance

called vanillin

Vanilla producing countries - Madagascar, Indonesia,

China, India and Mexico

In Sri Lanka very popular crop in home garden

systems

Main potential areas - Kegalle, Kandy, Matale, Nuwara-Eliya, Kurunegala and Gampaha districts

…..INTRODUCTION

The total export volume and export earnings (2011)

Processed vanilla - 0.2 Mt, 5.3 m rupees

Vanilla oil - 2.1 Mt, 3.3 million rupees

Propagation - either by stem cuttings or by tissue culture plants

Field planting - 1m cuttings from healthy, vigorous vanilla vine basically unflowered portion

Planting material requirement 2400 cuttings per hectare.

…. INTRODUCTION

limitation of planting materials

1 m long cutting contains about 12 nodes

Tissue culture technique – may be a best

alternative for production of planting materials

Advantages –

Mass production of plants within a short

period of time

production of disease free propagules

True to type plants

OBJECTIVES

To find a more suitable method for production

of planting material of vanilla

To compare tissue culture plants and plants

originated from stem cutting with traditional

method (one meter cutting)

METHODOLOGY

Location: Central Research Station - Matale

Net house experiment

Treatments:

T1 – Tissue culture plants

T2 – 2 nodal cuttings

T3 – 3 nodal cutting

T4 – 4 nodal cuttings

Replicates: 3

Number of plants per treatment: 25 cuttings

Experimental design - RCBD

…. METHODOLOGY

Polythene bag size – 30.4cm x 25.4cm

Standard potting mixture - top soil, coir dust,

sand and cow dung (1:1:1:1 ratio)

Growth Parameters collected:

Shoot length

Number of new leaves

Girth

Data were collected at monthly interval

Experimental period – 5 months

…. METHODOLOGY

Field experiment

Treatments:

T1 – tissue culture plants

T2 – 2 nodal rooted cuttings

T3 – 3 nodal rooted cuttings

T4 – 4 nodal rooted cuttings

T5 – 1m rooted cuttings (control)

Plants originated from the same treatments were evaluated in the field

Replicates: 3

Number of vines per treatment: 12

…. METHODOLOGY

Gliricidia sticks were used as live support

Gliricidia supports were planted 6 months

before vanilla planting

Spacing - 244cm x152.5cm (8’x5’)

Vine training - As single training system

Make 4 - 5 loops around support tree

…. METHODOLOGY

Data collection

Growth parameters collected

length of the vine

Number of leaves

Number of loops

Girth

Yield parameters collected

Number of flower clusters

Number of flowers

Number of pods developed

RESULTS AND DISCUSSION

Net house experiment

The percentage of success very high in cuttings as

well as tissue culture plants (>90%)

Statistically significant differences were observed in

shoot growth, girth in tissue culture plants and 2

nodal cuttings compared to 3 &4 nodal cuttings.

Both four nodal and three nodal cuttings showed

significantly higher shoot growth, number of new

leaves than two nodal cuttings and tissue culture

plants

…. RESULTS AND DISCUSSION

Establishment of vanilla cuttings under net house

conditions

After establishment of vanilla

cuttings and tissue culture

plants, each vine was tied to a

104.2 cm (31/2 feet) long

dead coffee stick.

Vines were trained along the

stick to prevent entangle of

vines

Establishment of vanilla cuttings and tissue culture plants

under net house condition after four months

Tissue culture

plants

2 nodal

cuttings

3 nodal cuttings 4 nodal

cuttings

…. RESULTS AND DISCUSSION

Growth parameters of different cutting types and tissue

culture plants after five months of establishment

Treatment Shoot growth

(cm)

No. of new

leaves

Girth

(mm)

Tissue culture

plants 8.6 c

3.3 b

2.0 d

2 nodal cuttings 41.5 b

5.4 b 4.1 c

3 nodal cuttings 70.2 a

8.0 a 5.1 b

4 nodal cuttings 72.5 a

8.6 a 6.1 a

LSD 5% 17.49 2.4 0.5

Means with the same letter in a column are not significantly different p<0.05

0

10

20

30

40

50

60

70

80

1M 2M 3M 4M

Sh

oo

t g

row

th (

cm

)

Duration (Months)

TC

2N

3N

4N

Comparison of shoot growth in different cutting types (2, 3

and 4 nodal and tissue culture plants (TC) with the time

…. RESULTS AND DISCUSSION

…. RESULTS AND DISCUSSION

Growth rate of tissue culture plants was

significantly lower than all other type of cuttings

Cuttings with higher length and higher number of

leaves show higher growth rate than the smaller

cuttings, might be due to more food reserves in

larger cuttings

Plants produced by tissue culture are very small

took long time to adopt in soil as compared to other

treatments, but plant survival is very high in tissue

culture plants.

After field

establishment of

vanilla rooted

cuttings and

tissue culture

plants

Experimental field - at initial stage

…. RESULTS AND DISCUSSION

…. RESULTS AND DISCUSSION

Field experiment

After 3 years vines from all the five types of

planting materials showed similar growth and no

significant differences among the treatments

(planting materials)

Flowering started in all five types of vines at the

same time

The percentage of plant that flowered varies

with the type of planting materials

Growth parameters of vines raised from different planting

materials in the field

Treatment New Shoot

Growth (cm)

Number of leaves Number of

loops

Girth (mm)

2nd

year

3rd

year

2nd

year

3rd year 2nd

year

3rd

year

2nd

year

3rd

year

Tissue

culture

Plants

184.9 a

400.7 a

20.4 b

23.7 b

0.9 a

1.0 a

17.8 b

19.5 a

2 nodal

316.2 b

357.6 a

33.0 ab

38.6 a

1.7 a

2.5 a

19.4 ab

21.7 a

3 nodal

313.0 b

355.5 a

49.5 a

46.3 a

1.2 a

1.5 a

20.8 ab

21.1 a

4 nodal

293.6 b

317.6 a

44.7 a

39.3 a

1.3 a

1.6 a

18.2 b

18.9 a

1m cutting

286.0 b

410.4 a

19.5 b

24.3 b

1.3 a

2.4 a

22.2 a

22.9 a Means with the same letter in a column are not significantly different p<0.05

….RESULTS AND DISCUSSION

The lowest percentage of plants that flowered was

observed in tissue culture plants (5.7 %) followed by

two nodal cuttings (6.7%) while the percentage of

plants that flowered were higher in three nodal

(30.6%), followed by 1m long cuttings (24.4%) and

four nodal cuttings(22.2%)

Vines with lowest number of pollinated flowers

were observed in tissue culture plants followed by

two nodal cuttings while number of pollinated

flowers were higher in 1 m long cuttings followed by

four nodal and three nodal cuttings

Number of pollinated flowers of different cutting types and tissue

culture plants

-50

0

50

100

150

200

250

300

Tissueculture

2 nodalcuttings

3 nodalcuttings

4 nodalcuttings

1 metercuttings

Nu

mb

er

of

po

llin

ate

d f

low

ers

Planting material

1st flowering

2ndflowering

3rd flowering

…. RESULTS AND DISCUSSION

In field three years after planting no significant

difference in growth observed among different

treatments. But at the flowering stage, higher

number of flowers were observed in three nodal,

four nodal and 1m long cuttings

CONCLUSIONS

Three nodal and four nodal cuttings of vanilla

could be used as planting materials as an

alternative to 1m long cuttings for vanilla

cultivation

Tissue culture plants did not perform well in

both nursery stage and field as compared to

the plants originated from stem cuttings

Growth and reproductive characteristics are

better when the plant originating from larger

cutting size

REFERENCES

Anonymous (2011): Performance and Accounts Reports, Department Export

Agriculture, Peradeniya, pp 54 – 55.

Gunasinghe, U. M. (2007): Vanilla Cultivation. Department of Export

Agriculture, Peradeniya, Sri Lanka.

Hartmann, H.T., Kester, D.E., Davies, F.T and Geneve, R.L. (1995): Biology

of propagation of cuttings. In: Plant propagation Principles and Practices,

Prentice Hall Inc., New Jersey, pp 288-289.

Juan Hernandez-Hernandez, (2011). Mexican vanilla production. Handbook

of Vanilla Science and Technology, pp 1-5 & 14- 16.

Price, B. (1980): Plant propagation. Granada Publishing, pp 15-87.

Rao, Y.S., Mathe, K.M., Lakshmanan, R., and Kumara, P.K. (2002): Tissue

culture and its Implications in Vanilla. Spice India 15: 8-11.

Smithy, M. (2003): The wonderlands of Vanilla. Features. http:www.

orchidsasia.com Accessed on 24th March, 2006.

Torres, K.C. (1989): Tissue culture Technique for horticultural crops.

Chapman and Hall. New York, pp 26-48.

Thrulinger, R.W. (1948): Vanilla culture in Puerto Rico, Federal Experiment

Station in Puerto Rico, USDA Experiment Station, pp 8-15.