Gamma Ray Application for Increasing Kemenyan Toba (Styrax

sumatrana) Seed Viability

Cut Rizlani Kholibrina

, Aswandi

and Arida Susilowati

Environment and Forestry Research and Development Institute of Aek Nauli. Jl. Raya Parapat Km 10,5. Simalungun,

North Sumatra

Faculty of Forestry, Universitas Sumatera Utara. Jl. Tridharma Ujung No.1 Kampus USU Medan, 20155

Keywords: Styrax, Seed, Germination, Viability, Gamma Ray.

Abstract: Kemenyan toba (Styrax sumatrana) is the identity trees for Tapanuli community in North Sumatra, Indonesia.

These non-timber forest products have been cultivated for generations. Over the past decade, the incense

productivity have been decreasing due to the lower interest of farmer to cultivate kemenyan. The another

reason also causing by longer germinate time so that the farmers difficulty obtaining high quality seedlings

for replanting the old unproductive trees. Accelerate germination time through improving seed viability and

genetic engineering were prospective ways for increasing kemenyan productivity. The objective of this research

was to determine the effectiveness of gamma irradiation techniques application for increasing the viability of Kemenyan toba seeds.

Randomized completely design, with 5 levels of radiation dose those were, control 0 Gy, 10 Gy, 20 Gy, 30

Gy, and 40 Gy with 4 replications was used in this research. The results showed that increasing the intensity of

irradiation shortens the germination time. The highest germination rate occurred on day 61 on the intensity of

irradiation 40 Gy whereas the control was day 197. The treatment of irradiation affected the germination.

However, increasing irradiation intensity decreasing the sprouting ability. On seeds without irradiation,

sprouting percentage reach an average of 83.8%. The germination rate was not different compared to the seeds

that received irradiation treatment with intensities of 10 and 20 Gy with sprout power of 75.0% and 57.5%. It

means that the low dose of gamma ray irradiation can be used to increase the viability and vigor of Kemenyan seeds.

1 INTRODUCTION

One of valuable non-timber forest product that has a

long history and become the main community

livelihood in Tapanuli region, North Sumatra is

Kemenyan rosin. Historically and economically this

commodity has been cultivated from Styrax spp trees

for a long time and is a major source of regional

income (Kholibrina et al., 2018). BPS Sumut (2018)

reported that incense production reaches 5,661.39

tons every year. It mean that if the price of incense at

the farmer level reaches Rp 200 thousand/kg, the

farmer will resulting around Rp1.2 trillion of income

every year from kemenyan forest management.

Although valuable comodity, the sustainability of

kemenyan production were constrain by some

problems. In the last decade, the population of

incense trees has declined due to logging and forest

conversion (Susilowati et al., 2018). The low

willingnes of farmer to replant their unproductive

trees also contribute the lower rosin production.

Furthermore, Styrax seed also takes a long time for

germinate. BPS North Sumatra (2018) states that

kemenyan rosin productivity has been decline from

6,060.89 tons/ha in 2008 to 5,661.39 tons/ha in 2017.

Therefore, it is necessary rapid effort to increase the

rosin productivity in North Sumatra, one of which by

improving seed quality.

The utilization of high quality seeds is the starting

point of a stand development and improving

kemenyan rosin quality. These efforts can be started

from the seed stage, through increasing seed viability

and vigor. Seed viability and vigor determine seed

quality both physically and physiologically. Seed

quality improvement can be conducted by gamma ray

irradiation techniques (Piri et al., 2011). Aplication

the irradiation techniques to improve vigor and seed

quality has been carried out on agricultural crops, but

in forest plant seeds are still limited (Iglesias-Andreu

et al., 2012). In trees species, the application of

gamma ray radiation at low doses can improve seed

germination and seedling growth (Iglesias-Andreu.,

Kholibrina, C., Aswandi, . and Susilowati, A.

Gamma Ray Application for Increasing Kemenyan Toba (Styrax sumatrana) Seed Viability.

DOI: 10.5220/0008387200200025

In Proceedings of the International Conference on Natural Resources and Technology (ICONART 2019), pages 20-25

ISBN: 978-989-758-404-6

2012; Akshatha et al., 2013). Chan & Lam (2002)

also reported that irradiation of papaya seeds with a

dose of 10 Gy increased the germination percentage

to 50%. Zanzibar & Witjaksono (2011) reported that

irradiation of Suren seeds (Toona sureni) using low

doses (5 Gy) able to increase the seedling growth rate.

The plant sensitivity to radiation treatment depends

on many factors, such as species or varieties, plant

parts and the radiation doses (Esnault 2010; de Micco

et al., 2011). The objective of this study was to

determine the effectiveness of gamma ray irradiation

techniques to increase the seed viability of Kemenyan

toba (Styrax sumatrana).

2 METHODS

The material of this research was kemenyan seeds,

and germination media. Kemenyan toba seeds for the

research was obtained from Humbang Hasundutan

district, North Sumatra Province. The gamma ray

irradiation dose used were 0 Gy (control), 10, 20, 30,

and 40 Gy. The Gamma irradiation was conducted at

the National Atomic Energy Agency (BATAN),

while seed viability and vigor testing were conducted

in greenhouse in Environment and Forestry Research

and Development Institute of Aek Nauli in Lake Toba

region, North Sumatra.

Randomized completely design with irradiation

dose treatments was used in this research. The treated

seeds was germinated in polybags 12x17cm in size

using sterilized mixture of soil sand media (1:1/v:v).

The number of seeds sown for five treatments with 4

replications and 20 observation units reached 400

seeds. Observation of germination is carried out every

day since being planted until no seeds germinate. The

observation variables include days of germination

which ae marked by the appearance of the radicles,

germination rate, seedling height, diameter, branches

and number of leaves. Data were analysed using

analysis of variance to determine variation between

treatments. If there were variations, the analysis was

continued with Duncan's Multiple Distance Test

(Duncan 's Multiple Range Test - DMRT).

3 RESULT AND DISCUSSION

The results of variance analysis for germination day,

germination rate, diameter and height’s seedlings,

branching and number of leaves on irradiated seeds

are shown in Appendix 1, Table 1 and Table 2.

Table 1: Analysis of variance for the observation variables

of duration and germination rate.

Treatments

Germination days

Germination rate

Control

197

83.75

10 Gy

102

(-48%)

75.00

(-10%)

20 Gy

(-65%)

57.50

(-31%)

30 Gy

(-67%)

25.63

(-69%)

40 Gy

(-69%)

4.15

(-96%)

The irradiation treatment affects the rate of

germination. Increasing the intensity of irradiation

shortens the germination time. Based on Table 1, the

normal seed (non treatment) starts the germination on

197

day. The shortest germination time occurred on

day 61 at the intensity of irradiation of 40 Gy, thus

reducing the germination time of 69% from normal

time. The 40 Gy dose also gives the highest seedling

high variable of 14.45 cm (Table 2). However, this

was not followed by an increase in germination rate

which decreased to 96%.

Table 2: Variance analysis for the seedling diameter, height,

branches and number of leaves.

Diameter

(mm)

Height*

Branches

Number of

leaves*

Control

157

2.3

10 Gy

103

(-35%)

2.6

abc

(-26%)

20 Gy

105

(-33%)

2.2

(-38%)

30 Gy

116

(-25%)

1.4

(-34%)

40 Gy

145

(-7.7%)

2.5

(5.33%)

Generally, application of gamma irradiation on

Kemenyan seeds provides two response those were:

supporting and inhibiting germination. The

irradiation treatment affects the speed of germination.

Increasing the intensity of irradiation will shortens

germination time. According to Zanzibar (2015), the

irradiation technique is an ionic process, when

ionizing radiation is absorbed into biological

material, the radiation will act directly on the critical

target cells or indirectly through the generation of

metabolites which can modify important cell

components. According to Luckey (1980) irradiation

at low doses can stimulate the physiological process

(radiostimulation) of plants through excitation or

known as hormesis. The influence of hormesis on

various agricultural crops species can provide a

positive or beneficial response (Luckey, 2003; Piri et

al., 2011).

The germination rate decreases with increasing

irradiation intensity. Based on Tukey HSD and

Duncan test, there were three groups of responses

(Table 1). On seeds without irradiation, percent

germinate reached 83.8%. The germination rate was

not different with another treatment (intensities of 10

Gamma Ray Application for Increasing Kemenyan Toba (Styrax sumatrana) Seed Viability

and 20 Gy with germination rate of 75.0% and

57.5%).

Increased doses of up to 40 Gy reducing sprouting

ability to 4.1% or decrease to 95.5% (Table 2).

Therefore, the dose for kemenyan seeds should be

lower than 40 Gy. Zanzibar (2015) states that the dose

level application and its effect on seed germination

varies for each species and genotype. But in general,

higher irradiation doses tend to inhibit germination.

Habba (1989) reported that an increase in irradiation

doses of up to 100 Gy increased seed germination

gradually, but then seed germination decreased with

increasing irradiation doses. A high irradiation dose

causes higher cell damage because the energy

released by gamma rays is quite large and penetrates

deeply. The amount of damage to cells cause’s lower

opportunities for survive (Hapsari, 2004). Wulandari

(2003) on Chrysanthemum found that increasing

gamma ray irradiation doses of 10, 15, 20 to 25 Gy

decreased the percentage of plant life.

The germination phase of all seeds shows a

natural response as well as germination of kemenyan

without irradiation (control). The new response can

be seen in the seedling phase which is marked by

changing the shape of the leaves to wavy to curly and

the large number of branches (Figure 1). Changes in

plant morphology are common and are most easily

seen from the irradiation of plants. The diversity due

to gamma ray irradiation is most commonly found in

leaf pinnate, both in terms of colour and shape. The

diversity of seedling morphology due to irradiation is

characterized by the occurrence of abnormalities or

malformations of plant organs. Hartati (2000) states

that irradiation treatment will cause cell damage or

inhibition of cell metabolism due to interference with

RNA synthesis so that the synthesis of enzymes

needed for growth is inhibited. This phenomenon

might be resulting enzyme to lose its function. The

irradiation treatment can cause enzymes that

stimulate growth to become inactive. Soeranto in

Herison (2008) states that the occurrence of

abnormalities in irradiated populations shows that

there have been changes at genomes level,

chromosomes, and DNA or genes that are very large

so that genetically controlled physiological processes

in plants become abnormal and cause variations in

new genetic variations. Abnormalities to plant

irradiation deaths are caused by the formation of free

radicals such as Ho, which are highly labile ions in

the reaction process due to irradiation. It resulting in

many collisions in various directions, which

consequently will make changes or mutations at the

DNA, cell and tissue levels and organs, even causing

death in plants. Abnormalities began to appear since

the leaves of the irradiated plants began to develop, at

47 HST. Abnormalities occur in the shape of leaves

in plants for all irradiation doses

Figure 1: The changes of the leaves shape and number of

branches.

Furthermore, irradiation affects the number of

leaves. The number of leaves in normal conditions

reaches an average of 10 sheets. The number of leaves

decreased after the seed was irradiated at an intensity

of 10 Gy to 8 sheets. But statistically the response of

the two treatments is the same. In seeds with

irradiation intensity of 20 and 30 Gy the number of

leaves decreases to 6 and 7 consecutive leaves. But

the increase in intensity to 40%, the number of leaves

increased to 11 sheets. Marcu et al (2012) on Lactuca

sativa plants, showed that the effective dosage for

increasing germination was not more than 30 Gy.

While at doses above 70 Gy, the vegetative part of

plant growth began decrease.

The irradiation treatment affects plant height. In

seeds without irradiation treatment, the response of

height reached 15.6 cm. At dose of 40 Gy the

response of the plants average height decreases to

14.45 cm. This high response is not statistically

different with the seeds with irradiation intensity 30

Gy. Conversely, irradiation treatment does not affect

stem diameter and number of branches.

ICONART 2019 - International Conference on Natural Resources and Technology

4 CONCLUSIONS

Our research point out that, increasing the intensity of

irradiation reducing the germination time from 197

days (without treatment) become 61 days (dose 40

Gy). Increasing irradiation intensity also decreasing

the sprouting ability from 83.8% (without treatment)

to 75.0% and 57.5% (dose 10 and 20 Gy). The

irradiation also affects the number of leaves. The

number of leaves was 10 sheets (without treatment)

but decreases after irradiated at an intensity of 10 Gy

to 8 sheets. The irradiation treatment also affects plant

height. But statistically the response of the two

treatments is the same. Conversely, irradiation

treatment does not affect stem diameter and number

of branches.

ACKNOWLEDGEMENTS

I would like to extend thanks to Wijaya Murti

Indriatama, Agriculture researcher on National

Atomic Energy Agency (PAIR BATAN).

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APPENDIX

ANOVA Germination days

Source

Sum of

Squares

Mean

Square

Sig.

Corrected

Model

52483.5

13120.9

11.69

.000

Intercept

195881.

195881.4

174.59

.000

Perlakuan

52483.5

13120.9

11.69

.000

Error

16829.0

1121.9

Total

265193.

Corrected

Total

69312.6

a. R Squared = .757 (Adjusted R Squared = .692)

Post Hoc Tests of Germination days

Treatment

Subset

Tukey

HSD

a,b

61.2250

65.9250

68.4500

101.9500

197.2750

Sig.

.452

1.000

Duncan

a,b

61.2250

65.9250

68.4500

101.9500

197.2750

Sig.

.133

1.000

The error term is Mean Square(Error) = 1121.936.

a. Uses Harmonic Mean Sample Size = 4.000.

b. Alpha = 0.05.

ANOVA of Germination rate

Source

Sum of

Squares

Mean

Square

Sig.

Corrected

Model

21137.6

5284.4

7.7

.001

Intercept

52439.0

76.4

.000

Perlakuan

21137.6

5284.4

7.7

.001

Error

10288.5

685.9

Total

83865.2

Corrected

Total

31426.2

a. R Squared = .673 (Adjusted R Squared = .585)

Post Hoc Tests of Germination rate

Subset

Tukey

HSD

a,b

4.1500

25.6250

57.5000

75.0000

83.7500

Sig.

.073

.107

.331

Duncan

a,b

4.1500

25.6250

57.5000

75.0000

83.7500

Sig.

.264

.106

.082

Means for groups in homogeneous subsets are displayed.

Based on observed means.

The error term is Mean Square(Error) = 685.900.

a. Uses Harmonic Mean Sample Size = 4.000.

b. Alpha = 0.05.

ANOVA of Diameter

Source

Sum of

Squares

Mean

Square

Sig.

Corrected

Model

1.228

.307

1.398

.282

Intercept

198.671

905.224

.000

Perlakuan

1.228

.307

1.398

.282

Error

3.292

.219

Total

203.190

Corrected

Total

4.520

a. R Squared = .272 (Adjusted R Squared = .077)

ICONART 2019 - International Conference on Natural Resources and Technology

ANOVA of Height

Source

Sum of

Squares

Mean

Square

Sig.

Corrected

Model

93.973

23.493

3.220

.043

Intercept

3125.625

428.430

.000

Perlakuan

93.973

23.493

3.220

.043

Error

109.433

7.296

Total

3329.031

Corrected

Total

203.406

a. R Squared = .462 (Adjusted R Squared = .319)

Post Hoc Tests of Height

Subset

Tukey

HSD

a,b

10.2500

10.5250

11.6250

14.4500

15.6562

Sig.

.080

Duncan

a,b

10.2500

10.5250

11.6250

14.4500

15.6562

Sig.

.060

.063

The error term is Mean Square(Error) = 7.296.

a. Uses Harmonic Mean Sample Size = 4.000.

b. Alpha = 0.05.

Table ANOVA of Number of Leaves

Source

Sum of

Squares

Mean

Square

Sig.

Corrected

Model

67.022

16.755

3.271

.041

Intercept

1409.521

275.127

.000

Perlakuan

67.022

16.756

3.271

.041

Error

76.848

5.123

Total

1553.390

Corrected

Total

143.869

a. R Squared = .466 (Adjusted R Squared = .323)

Table Post Hoc Tests of Number of Leaves

Subset

Tukey

HSD

a,b

6.3750

6.8500

7.6000

10.3000

10.8500

Sig.

.085

Duncan

a,b

6.3750

6.8500

7.6000

10.3000

10.8500

Sig.

.480

.058

.072

The error term is Mean Square(Error) = 5.123.

a. Uses Harmonic Mean Sample Size = 4.000.

b. Alpha = 0.05.

ANOVA of Number of Branch

Source

Sum of

Squares

Mean

Square

Sig.

Corrected

Model

3.282

.821

1.485

.256

Intercept

96.141

173.972

.000

Perlakuan

3.282

.821

1.485

.256

Error

8.289

.553

Total

107.713

Corrected

Total

11.571

a. R Squared = .284 (Adjusted R Squared = .093)

Gamma Ray Application for Increasing Kemenyan Toba (Styrax sumatrana) Seed Viability