The Effects of Colchicine Concentration and Length of Immersion on

Cutting Growth of Patchouli (Pogostemon cablin Benth)

Zuyasna

, Andre

and Siti Hafsah

Faculty of Agriculture, Syiah Kuala University, Jl.T.H.Krueng Kalee, Banda Aceh, Indonesia

Pidie Regency Agriculture and Food Service, Sigli, Indonesia

Keywords: mutation, diversity, colchicine, immersion, patchouli

Abstract: The aims of the study were to determine the concentration of colchicine and the best immersion length for the

growth of patchouli cuttings. This study used a Randomized Block Design (RBD) 4 x 4 factorial pattern with

5 replications, the factors tested were: Colchicine concentration consisted of 4 levels (C0: Without Colchicine,

C1: 0.25% Colchicine, C2: 0.50% Colchicine and C3: 0.75% Colchicine) and immersion length consists of 4

levels (R1: 2 hours, R2: 4 hours, R3: 6 hours, and R4: 8 hours). This research conducted in Sigli - Pidie

Regency, Aceh-Indonesia from May to July 2016. The colchicine concentration affected the height, leaf area,

and number of patchouli branches. Length of immersion give a different response. There was an interaction

between the concentrations of colchicine and length of immersion in plant height, but there was no interaction

on leaf area and number of patchouli branches.

1 INTRODUCTION

Pogostemon cablin Benth is plant that produce the

essential oil and has high economic value in the

world. This plant is an important crop in Indonesia

because it can contribute a high foreign exchange to

the country (Hariyani et al., 2015). This essential oil

is one of the most important naturally occurring

perfumery raw materials because of its characteristic

woody fragrance and ﬁxative properties by which the

scent is ﬁxed and make it last longer on the skin.

Patchouli essential oil produced from the distillation

process of the patchouli leaves. Pogostemon cablin

(P.cablin; common known as Patchouli) originated

from southeast Asia is cultivated extensively in

Indonesia, Philippines, Malaysia, China, and Brazil

(Miyazawa et al., 2000; Singh et al., 2002; Wu et al.,

2008). The aerial part of P.cablin has been used for

the treatment of the common cold, headache, fever,

vomiting, indigestion and diarrhea as well as an

antifungal agent in the medicinal materials of China

and its surrounding regions (

Board of Pharmacopoeia of

P. R. China). It is an herbaceous perennial plant with

oil glands producing an essential oil (patchouli oil),

which is commonly used to give a base and lasting

character to a fragrance in the perfume industry.

Due to its uses in perfumery, the demand of

patchouli oil is increasing dramatically in the world.

Therefore, available preparations of the patchouli oil

products may differ significantly in quality depending

on a number of factors such as the plant varieties,

tissues or organs used, harvesting time (different

developmental stages of the plant) and the different

and poorly controlled analysis conditions (Bergonzi

et al., 2001). In addition, the geographic location was

an important factor affecting the chemical

composition and developmental process of the

medicinal plant. The plant growth progress and

chemical characterization varied under different

environmental conditions and cultivation locations.

Indonesia has three types of patchouli namely

Pogostemon cablin Benth, Pogostemon heyneatus

Benth, and Pogostemon hortensis. Patchouli plants

are vegetative propagated by cuttings, because

propagation through seeds is not possible, this is

because patchouli plants do not have flowers even

under the photoperiodical control (Hefendehl and

Murray, 1979), so it does not allow pollination and

fertilization. Therefore, the diversity of patchouli

plants is very narrow and undeveloped, so it needs

efforts in increasing plant diversity by mutation

approach. A possible alternative is to produce a

mutant trait in order to have a new clone.

Zuyasna, ., Andre, . and Hafsah, S.

The Effects of Colchicine Concentration and Length of Immersion on Cutting Growth of Patchouli (Pogostemon cablin Benth).

DOI: 10.5220/0009973001590164

In Proceedings of the 2nd International Conference of Essential Oils (ICEO 2019), pages 159-164

ISBN: 978-989-758-456-5

159

Mutation induction is one of the non-

conventional plant breeding methods that aims to

increase the genetic diversity of a plant. Mutations are

changes in genetic material in living things that occur

suddenly and randomly inherited. Mutations that

occur inherited and can return to normal (epigenetic).

Mutations can occur naturally or intentionally

induced for certain purposes for genetic improvement

of plants. Natural mutations can occur due to the

presence of sunlight, as well as electrical energy such

as lightning. Artificial mutations for plant breeding is

by giving mutagens. There are two groups of

mutagens that used to get mutants, physical mutagens

and chemical mutagens. Physical mutagens are x-

rays, gamma rays and ultra violet rays. While

Chemical mutagens are Ethyl Methane Sulfonate,

Diethyl sulphate, Ethyl Amin and colchicine.

Colchicine is a toxic and carcinogenic alkaloid

obtained from the extract of the Colchium autumnale

plant and various other members of the Colchicaceae

tribe (Eka et al., 2014). Colchicine is applied to the

part of the plant that is actively dividing at a

vegetative growth point so that it can inhibit the

metaphase stage. Giving colchicine known to affect

plant growth, such as producing changes in plant

morphology, decreasing plant height, stem

circumference diameter, leaf area, number of crop

flowers and number of plant capsules and increasing

the flowering age of a plant. The addition of

colchicine by dropping at the growing point affected

the plant height and diameter of the lower stem

circumference, then the leaf area became narrower,

the flowering period was longer, but the percentage

of plants that produced higher seeds than using the

immersion technique at the tip of the sprouts (Sri et

al., 1999).

In this study, we conducted the use of colchicine

with several concentrations and length of immersion

to the patchouli of Lhokseumawe var. in an effort to

increase the genetic diversity and productivity of

patchouli plants.

2 MATERIALS AND METHODS

This research conducted in Sigli, Pidie Regency,

Aceh-Indonesia. This research took place from May

to July 2016. This study used 4 x 4 factorial

randomized block design (RBD). The factors that

were tried were: The concentration of colchicine

consisted of 4 levels (C0: No Colchicine; C1: 0.25%

Colchicine; C2: 0.50% Colchicine, and C3: 0.75%

Colchicine). The immersion length consists of 4

levels (R1: 2 hours; R2: 4 hours; R3: 6 hours, and R4:

8 hours). Thus, there were 16 treatment combinations

with 5 replications, so this study consisted of 80

experimental units. The planting media used in this

study was top soil mixed with husk and compost with

ratio of 1: 1:1, which then filled in a 17 × 23 cm

polybag.

2.1 Preparation of Colchicine Solution

Colchicine solution made as much as 1 L for each

concentration. Colchicine with a level of 0.25%

obtained by weighing 2.5 mg of colchicine and then

put into a measuring cup and dissolved with distilled

water up to 1 L. For 0.50 and 0.75% colchicine

solution, 5.0 and 7.5 mg of colchicine weighed and

dissolved with distilled water up to 1 L.

2.2 Preparation for Planting Material

(Patchouli Cuttings)

The planting material cleaned with distil water. Then

immersed in a colchicine solution according to the

treatment level of concentration and soaking time.

Then rinse with distilled water.

2.3 Planting

The planting material that has been prepared directly

planted on the media according to the treatment.

2.4 Maintenance

Plant watered every day and when needed the areal

plant cleaned from weeds.

2.5 Observation

The factors observed in this study were: a) Plant

Height (cm), plant height measured at 30, 60 and 90

DAP (day after planting). b) Leaf area (cm

), leaf area

was observed at 30, 60, and 90 DAP, using millimetre

paper. c) Number of branches, the number of

branches calculated from the base of the stem to the

growing point at 30, 60, and 90 DAP.

3 RESULT AND DISCUSSION

3.1 Plant Height

Results on the analysis of variance showed that the

concentration of colchicine and length of immersion

did not significantly affect the average height of

ICEO 2019 - 2nd International Conference of Essential Oil Indonesia

160

patchouli seedlings at the age of 30 days after planting

(DAP), but had a very significant effect at age 60, and

90 DAP.

Plant height is an indicator of growth that is

easiest to observe and used to observe the effects of

environmental influences and the treatments applied

(Sitompul and Guritno, 1995). Table 1 shows that in

the plant height on 30 DAP, the highest value was

found in the control treatment with a value of 2.41 cm

but not significantly different from other treatments.

At the age of plants 60 and 90 DAP, the highest value

was found also in the control treatment with a value

of 4.70 cm at 60 DAP and 10.56 cm at 90 DAP, which

was very significantly different from the treatment of

colchicine.

Table 1: Average patchouli plant height on 30, 60 and 90 DAP due to immersion with colchicine

Treatment

Average Plant Height (cm)

30 DAP 60 DAP 90 DAP

Colchicine Concentration

C0 (control) 2,41 4,70d 10,56d

C1 (0,25%) 2,33 4,29c 8,89c

C2 (0,50%) 2,23 3,51b 7,15b

C3 (0,75%) 2,11 2,91a 6,55a

LSD

0,05

- 0,33 0,33

Immersion Length

R1 (2 hour) 2,33 3,96 8,59b

R2 (4 hour) 2,35 3,94 8,22a

R3 (6 hour) 2,23 3,87 8,21a

R4 (8 hour) 2,17 3,64 8,11a

LSD

0,05

- - 0,33

Note: numbers followed by different letters on the same line are significantly different at 0.05 LSD

This is can be assumed that colchicine

succeeded in causing the plant cell size become larger

but plant height becomes lower, so that high

concentrations of colchicine can inhibit the growth of

patchouli. Permadi et al. (1991) mention that the

greater chance of inhibition of plant height followed

by the higher concentration of colchicine, and

Honkanen et al., (1992) in his research also found that

colchicine affects the growth of gerbera plants at a

high level of concentration. Mihu et al. (1989) stated

that 0.2% colchicine concentration showed a decrease

in shoot height in cabbage (Brassica oleraceae)

plants.

Inhibition of plant height is not only influenced

by the concentration of colchicine, but based on the

results of the F test on the analysis of variance shows

that length of immersion also has a very significant

effect on the average height of patchouli at the age of

90 DAP. The effect of length of immersion of

colchicine causes stunted plant height growth when

compared to non-treatment with colchicine. This can

occur due to colchicine dissolved in stem cells

affecting cell division, so the process becomes slower

when compared to cells in normal shoots. This also in

accordance with Permatasari (2007) research that the

treatment with the longest immersion showed the

lowest average height of Hibiscus rebaudiana plant.

Sri et al., (1999) reported that the technique and

immersion with colchicine at the level of 0.05% at the

point of growth of Hibiscus sp resulted in a decrease

in plant height.

3.2 Leaf Area (cm

)

The analysis of variance showed that the

concentration of colchicine and length of immersion

did not significantly affect the average leaf area at 30

DAP, but had a very significant effect at age 60 and

90 DAP Table 2 showed that the highest leaf area

obtained in the control compared to the leaf area in

the treatment immerse with colchicine. Leaf area is a

growth parameter that can determine the rate of

photosynthesis per plant unit (Sitompul and Guritno,

1995). Leaf growth is very important because it will

affect the fresh weight and dry weight produced;

especially the leaves are an important yield

component for patchouli plants. The results of this

study indicate that the treatment of colchicine causes

the leaf area sizes reduced compared to control plants

with have larger of area leaf sizes.

Table 2 also shows that in the plant height at 30

DAP there were no significant differences with other

treatments. However, there were very significant

differences in the observations of 60 and 90 DAP

from the concentration of colchicine and immersion

length, the interaction between the two factors had no

significant effect on leaf area. The results of this study

indicate that the colchicine has an effect on

The Effects of Colchicine Concentration and Length of Immersion on Cutting Growth of Patchouli (Pogostemon cablin Benth)

161

decreasing leaf area, where the higher the

concentration of colchicine given the narrower the

leaf area size. This maybe accordance with the

process of mitosis that occurs in disturbed cells due

to colchicine, which has toxic properties. Herman et

al., (2013) stated that phenotype changes due to

colchicine treatment not only had an impact on

changes in the number and size that were greater due

to colchicine treatment than its control, but also had

an impact on the narrowing of leaf area size.

Table 2: Rata-rata luas daun pada umur 30, 60 dan 90 DAP

akibat pemberian colchisine dan immersion length.

Treatment

Average leaf area (cm

)

30 DAP 60 DAP 90 DAP

Colchicine Concentration

C0 (Control) 9.47 19.41c 5.68c

C1 (0,25%) 9.43 18.96c 24.95c

C2 (0,50%) 8.99 16.51a 20.81b

C3 (0,75%) 8.55 13.68a 17.08a

BNJ

0,05

- 1.06 0.97

Immersion length

R1 (2 Jam) 9.52 18.63b 23.98c

R2 (4 Jam) 9.13 16.86a 21.88b

R3 (6 Jam) 8.88 16.95a 21.91b

R4 (8 Jam) 8.91 16.11a 20.75a

BNJ

0,05

- 1.06 0.97

Note: numbers followed by different letters on the same line

are significantly different at 0.05 LSD

The small size of plant leaves due to the

treatment of colchicine is caused by stress due to the

concentration and duration of immerse on colchicine,

so that the process of cell division is hampered due to

the colchicine which causes the primordial stage of

leaf formation to slow development (Haryanti et al.,

2009). In accordance with the results of Ajijah and

Bermawie's research on onion (2003), reported that

plants treated with colchicine can show the effect of

physiological damage, so that it can inhibit plant

growth, the effect of physiological damage seen in

leaf circumference size. The higher the concentration

of colchicine, the greater the effect of depression

(Permadi et al., 1991). The results of this study are

consistent with the results of the study of Ramesh et

al., (2011) who reported that mulberry plants soaked

in colchicine with concentrations of 0.1 to 0.3% had

smaller leaf area than controls.

In addition to the concentration of colchicine,

the immersion length factor also has a significant

effect on leaf area size. Two-hour immersion has the

largest leaf size, with a value of 18.63 at 30 DAP and

23.98 at 90 DAP. While the treatment with an eight-

hour immersion has the smallest leaf size, with a

value of 16.11 at 30 DAP and 20.75 at 90 DAP. It can

be said that the length of immersion length will result

in negative effects such as many damaged cells, thus

affecting the formation of a perfect leaf area.

According to Roberts, and Watson (2004) in

Anggraito Y. U (2004) states that the treatment time

is too long, then colchicine will show a negative

effect because cell degradation has occurred. The

results of this study are also consistent with the results

of the study of Yudia (2012), who reported that the

longest immersion treatment showed a trend that was

different from other immersion treatments. The

longest soaking with 0.02% colchicine solution

causes a decrease in leaf area size, but the number of

leaves is increasing.

3.3 Number of Branches

The analysis of variance showed that the

concentration of colchicine and immersion length did

not significantly affect the average number of

patchouli at the age of 30 DAP. Nevertheless, had a

very significant effect on the age of 60, and 90 DAP

(Table 3). At the age of 30 DAP the number of

branches did not have a significant difference due to

the treatment of colchicine and the immersion length.

However, at the age of 60 and 90 DAP there were

significant differences in the number of branches due

to the treatment of colchicine. The highest number of

branches was in the treatment of 75% colchicine

concentration with a value of 17.20 at the age of 60

DAP and 23.35 at the age of 90 DAP, compared to

the lowest in the control treatment with values 12.95

and 18 at the age of 60 and 90 DAP. It seems that

colchicine has an active role in increasing the number

of branches stimulate vegetative growth of plants.

The growth of branches for patchouli plants has a

positive effect on the yield produced by plants; it that

the better the growth of branches, the more likely the

growth of leaves will grow. The leaves are the main

target organ in patchouli as a producer of essential

oils.

The influence of colchicine on increasing the

number of branches shows that the concentration of

colchicine at the level of 75% is able to encourage

plants to induce number of branches / buds to grow

more. The results of this study are consistent with the

research of Haryanti et al., (2009) which states that

the dose of colchicine 0.20% affects the growth of

green bean plant cells. Plants experience an increase

in metabolic activity that stimulates branch growth

more than lower doses.

An increase in the number of branches indicates

that the application of colchicine may affect the

activity of genes that stimulate the activity of

ICEO 2019 - 2nd International Conference of Essential Oil Indonesia

162

hormones such as gibberellins, cytokinins, or inhibit

the production of auxins. Other research results state

that the application of colchicine can stimulate bud

induction and growth of buds of the Colophospermum

mopane plant, as well as stimulate growth in the

number of branches of tomato plants (Adelanwa et

al., 2011 in Sutrisno and Heru, 2014).

The immersion length factor also significantly

affected the number of branches at the age of 90 DAP,

where two-hour immersion had the fewest number of

branches, with a value of 19.50 and the highest was

found in the treatment with an eight-hour immersion

having a number of branches, with a value of 20, 90

at the age of 90 DAP. It is said that the length of

soaking time also has a certain optimal range to

produce the number of branches. Where the results of

Lina's research in 2010 showed that there was a very

significant effect on the number of new shoots due to

the duration of immersion, where the duration of

soaking with colchicine for 48 and 72 hours

decreased the number of new shoots, while the 24-

hour immersion treatment showed an increase in the

number of shoots. Lina (2010) states that the growth

of new shoots is inhibited because it is influenced by

the length of soaking time with colchicine, while the

results show that the 24, 48 and 72-hour immersion

treatment shows more shoot growth compared to

other immersion lengths.

Table 3: Average number of branches at 30, 60 and DAP

due to treatment with colchisine dan immersion length.

Treatment

Average number of branches

30 DAP 60 DAP 90 DAP

Colchicine Concentration

C0 (Control) 6.40 12.95a 18.00a

C1 (0,25%) 6.50 10.05a 18.75b

C2 (0,50%) 6.75 14.90b 21.00c

C3 (0,75%) 6.90 17.20c 23.35d

LSD

0,05

- 1.05 0.73

Immersion length

R1 (2 Jam) 6.50 14.15 19.50a

R2 (4 Jam) 6.50 14.30 20.15ab

R3 (6 Jam) 6.60 14.45 20.55b

R4 (8 Jam) 6.95 15.20 20.90b

LSD

0,05

- - 0.73

Note: numbers followed by different letters on the same line

are significantly different at 0.05 LSD

3.4 Interaction

The analysis of variance also showed that there was

an interaction between the concentration of

colchicine and immersion length on plant height at

the age of 90 DAP, but there were no interactions on

the measurement of leaf area and number of branches.

The average value of plant height on patchouli

seedlings aged 90 DAP between colchicine

concentration and immersion length on the growth of

patchouli as shows in Table 4.

There are interactions that cause a decrease in

the height of patchouli plants that given colchicine

and immersion length, so that the tendency in

treatment with colchicine has the lowest average

height of plants. This is probably due to by too high

concentrations of colchicine or immerse for too long.

According to Suryo (1995), plants will show negative

effects such as the number of damaged cells, stunted

growth, even causing the death of plants due to the

concentration of colchicine that is too high, or too

long immerse.

Research on patchouli plants by Mariska and

Lestari (2003) shows that immerse colchicine for too

long will reduce the mass of cells that can regenerate.

The highest percentage of regeneration is by soaking

colchicine for 1 day and the lowest by soaking for 7

days. Based on the results of Permatasari's research

(2007), it was reported that there was a decrease in

the average height of the Stevia rebaudiana bud in the

longest soaking colchicine treatment with a

concentration of 0.02% colchicine. This ensures that

the effect of giving colchicine treatment with the

length of soaking time provides greater opportunities

for colchicine dissolved into plant tissue. Lina (2010)

used of colchicine, it was able to suppress the average

height of shoots.

Table 4: Average patchouli plant height on 90 DAP

Colchicine

Concentration

Immersion length

LSD

0.05

(2hr)

(4hr)

(6hr)

(8hr)

C0 (Control)

10.84

9.94

10.68

Cab

10.76

Cab

0.88

C1 (0.25%)

9.32

8.9

8.8

8.52

C2 (0.50%)

7.28

7.2

7.12

7.00

C3 (0.75%)

6.92

6.84

6.26

6.16

4 CONCLUSIONS

The concentration of colchicine has no effect on plant

height, leaf area and the number of patchouli

seedlings at the age of 30 DAP, but there are

significant differences at age 60, and 90 DAP.

Immersion length does not affect plant height and

number of branches at 30 and 60 DAP, however there

is a significant difference at 90 DAP. Immersion

length has no effect on leaf area at 30 DAP, but there

The Effects of Colchicine Concentration and Length of Immersion on Cutting Growth of Patchouli (Pogostemon cablin Benth)

163

are significant differences at 60 and 90 DAP. There is

an interaction between the concentration of

colchicine and immersion length on plant height at

the age of 90 DAP.

REFERENCES

Ajijah,N. dan N. Bermawie. 2003. Pengaruh Kolkisin

Terhadap Pertumbuhan dan Produksi Dua Tipe

Kencur (Kaempferia galanga Linn). Buletin

Tanaman Rempah dan Obat 16 (1):46-55.

Anggraito, Y. U. 2004. Identifikasi berat, diameter, dan

tebal daging buah melon (Cucumis Melo, L.)

kultivar action 434 tetraploid akibat perlakuan

kolkisin. Berk. Hayati. 10:37–42.

Board of Pharmacopoeia of P. R. China (ed.),

“Pharmacopoeia of the People’s Republic of

China,” Chinese Edition 2010, Part I, Chemical

Industry Press,Beijing, 2010, pp. 42.

Eka J S, Bayu E S, Hasyim H. 2014. Pengaruh

concentration kolkhisin terhadap pertumbuhan dan

produksi kacang hijau (Vigna radiata L.) Program

Studi Agroekoteknologi, Fakultas Pertanian USU,

Medan. Jurnal Online Agroekoteknologi . ISSN

No. 2337- 6597. 2 (3):1238- 1244.

Evi N A, Respatijarti dan Sugiharto A N. 2016. Pengaruh

pemberian colchisine terhadap penampilan fenotip

Galur inbrida jagung pakan (Zea mays L.) pada fase

pertumbuhan Vegetatif. Jurnal Produksi Tanaman,

Universitas Brawijaya. Jawa Timur. 4 (5): 370-377

Hariyani, E. Widaryanto, N. Herlina. 2015. Pengaruh umur

panen terhadap rendemen dan kualitas minyak atsiri

tanaman nilam (Pogostemon cablin Benth.). Jurnal

Produksi Tanaman. 3 (3) : 205-211.

Haryanti, S., R. B. Hastuti, N. Setiari and A. Banowo. 2009.

The influence of colchicine to grow, metaphase cell

size and protein content of green beans plant (Vigna

radiata (L) Wilczek). Jurnal Penelitian Sains &

Teknologi 10(2):112-120.

Herman, Irma Natalina M dan Dewi Indriyani Roslim.

2013. Pengaruh Mutagen Kolkisin Pada Biji kacang

Hijau (Vigna radiata L.) Terhadap Jumlah

Kromosom dan Pertumbuhan. Jurusan Biologi

FMIPA Universitas Riau. Pekanbaru. J. BioETI. :

13-20.

Honkanen, J., A. Aapola, P. Seppanen, T. Tormala, J. C.

Wit, H. F. Esendam, L. J. M. Stravers, and J. C. De-

Wit. 1992. Production of doubled haploid Gerbera

clones. Acta Hortic.300: 341, 346

Lina, N .2010. Induksi mutasi kromosom dengan colchisine

pada Anthurium wave of love (Anthurium

plowmanii Croat.) secara in vitro. Skripsi.

Departemen Agronomi dan Hortikultura Fakultas

Pertanian Institut Pertanian Bogor. Bogor.

Mariska, I. dan E.G. Lestari. 2003. Pemanfaatan kultur in

vitro untuk meningkatkan keragaman genetik

tanaman nilam. Jurnal Litbang Pertanian 22(2):64-

69.

Mihu, G., N. Munteanu, and V. Timofte. 1989. Aspect of

some phenotypic changes induced by kolkisin in

cabbage. Cercetari Agronomice in Moldova.

22(4):85-93.

Mirzada, C. D. 1994. Pengaruh Beberapa Taraf BAP dan

IBA terhadapPerbanyakan Calla Lily secara In

Vitro. Skripsi. Program Sarjana, InstitutPertanian

Bogor. Bogor. 55 hal.

Miyazawa, M., Okuno, Y., Nakamura, S., Kosaka, H.,

2000. Antimutagenic activity of flavonoids from

Pogostemon cablin. J.Agric. Food Chem.48, 642-

647.

Ningsih E. M. N., Nugroho Y. A., dan Trianitasari, 2010.

Pertumbuhan Stek Nilam (pogostemon cablin,

benth) Pada Berbagai Komposisi Media Tumbuh

dan Dosis Penyiraman Limbah Air Kelapa.Jurnal

Agrika 4 (1): 37-47

Permadi, A. H., R. Cahyani, dan S. Syarif. 1991. Cara

pembelahan umbi, lama perendaman, dan

konsentrasi kolkisin pada ploidisasi bawang merah

Jurnal Pemuliaan Indonesia. Zuriat. Universitas

Padjadjaran. ISSN : 265-6261. 2 (2):36-41

Permatasari, D. 2007. Evaluasi Keragaman Fenotipe

Tanaman Stevia (Steviarebaudiana Bertoni M)

Klon Zweereners Hasil Mutasi Kromosom

denganKolkisin. Skripsi. Program Sarjana, Institut

Pertanian Bogor. Bogor.44 hal.

Ramesh, H. L., V. N. Y. Murthy and M. Munirajappa. 2011.

Colchicine induced morphological variation

mulberry variety M5. The Bioscan 6(1):115-118.

Singh, M., Sharma, S., Ramesh, S., 2002. Herbage, oil yield

and oil quality of patchouli [Pogostemon cablin

(Blanco) Benth.] influenced by irrigation, organic

mulch and nitrogen application in semi-arid tropical

climate. Ind. Crop Prod.16, 101-107.

Sitompul S.M. dan B. Guritno. 1995. Analisis Pertumbuhan

Tanaman. Gajah Mada University Press.

Yogyakarta.Hal 113-114.

Sri, R., Sudjindro, dan Basuki. 1999. Penggunaan

Colchicine dalam Penggandaan Kromosom Hasil

Hibridisasi Interspesifik pada Hibiscus sp. Untuk

Mengatasi Sterilitas F1. Tesis. Program

Pascasarjana Universitas Brawijaya. Malang

Suryo. 1995. Sitogenetika. Gajah Mada University Press.

Yogyakarta. 446 hal. Suryo. 1995. Sitogenetika.

Gajah Mada University Press. Yogyakarta. 446 hal.

Sutrino dan Heru. 2014. Keragaan Dua Varietas Kedelai

Pada Enam Concentration Colchisine. Balai

Penelitian Tanaman Aneka Kacang dan Umbi.

Malang.

Wu, Y.G., Guo, Q.S., Zheng, H.Q., Studies on residuals of

organochlorine pesticides and heavy metals in soil of

planting base and Pogostemon cablin. China J.

Chin.Materia Med.33(2008) 1528-1532.

Yudia P A. 2012. Induksi mutasi melalui penggandaan

kromosom nilam varietas sidikalang (Pogostemon

cablin Benth.) dengan colchisine secara in vitro.

Skripsi. Departemen Agronomi dan Hortikultura

Fakultas Pertanian Institut Pertanian Bogor. Bogor.

ICEO 2019 - 2nd International Conference of Essential Oil Indonesia

164