239

Journal of Medicinal and Aromatic Plant Sciences 32 (3) (2010) 239-244

ABSTRACT

Experiments were conducted to evaluate the growing behaviorof stem cutting of Coleus forskohlii Briq. Percentage of cuttingssprouted, nodes sprouted and survival were recorded to assessthe effect of cutting length, diameter and pretreatment withexogenous hormones viz., indole 3 acetic acid, -naphthaleneacetic acid and indole 3 butyric acid. The data revealed thesignificant effect of these factors on sprouting and survival ofC. forskohlii stem cuttings. Maximum sprouting, node sproutingand survival were recorded when cuttings were treated withIBA @ 100 ppm concentration as liquid and in powderformulation irrespective of diameter and length comparablewith control and water dip treatment. Powder formulations ofall hormones were found to be more effective than liquidformulations in relation to sprouting and survival of cuttings.Sprouting and survival of thin cuttings of four nodal lengthwere highest compared to other treatments.

Key words: Coleus forskohlii, sprouting, hormones, stem cutting

INTRODUCTION

Coleus forskohlii Birq., an indigenous medicinal herbwith tuberous root has been recorded to produce a root drug.In Ayurvedic Materia Medica under the Sanskrit name theplant was refered as Makandi and Mayani [13]. It is widelyused in various disorders viz. antiaging, antioxidant and aremedy for heart, abdominal and respiratory disorders [2, 6]. Itgrows on dry, barren hills and is cultivated in India for itsroots, which contains the diterpenoid ‘forskohlin’, thepresence of which is exclusive to this particular species [1,14]. The pharmaceutical industry is largely dependent uponwild populations for supply of plant material for forskolin. Infact, it is now listed as one of the plant species in Indiavulnerable to extinction due to indiscriminate collection ofthe plant from forests [15]. Thus, it is necessary to developmethods for the conservation of this threatened species whichis endangered and can be propagated in vitro.

Author for correspondence; E mail: rkstiwari@rediffmail.com

Sprouting and survival of Coleus forskohlii stem cuttings by hormonal pre-treatmentRAJ KUMAR SINGH TIWARI* AND KUNTAL DAS

Department of Plant Pathology, T.C.B. College of Agriculture and Research Station (I.G.K.V.), Bilaspur,Chhattisgarh, India.

Received 5th January, 2010

This herbaceous species is not only hard to propagatesexually but also show complexities and show undesirablecharacters if propagated through sexual means. Vegetativepropagation therefore, is the most vital and sole method toreproduce these plants species conserving the innate desirablecharacters [12]. Establishment and growth rate of the cuttingdepends upon many factors like seasonal and age variations,portion and diameter of branch, growing media, moisture level,nutrient status and temperature etc. [11]. In addition, the useof plant growth regulatory hormones - ‘auxins’, plays a vitalrole in influencing the important phases of plant growth anddevelopment.

Auxins are a class of plant growth substances oftencalled phytohormones or plant hormones. Among the auxinsIndole 3 Acetic Acid (IAA), á- Naphthalene Acetic Acid (NAA)and Indole 3 Butyric Acid (IBA) are typically the principalauxins used for rooting and sprouting of cuttings and majorityof plant species are responsive for rooting [7, 10]. Thesechemicals are available in commercial preparations, dispersedin concentrated liquid (liquid formulation), or in talc (powderformulation), that can be diluted with water (aqueous solution)to proper strength.

The present investigation was conducted in view of thevegetative propagation of Coleus forskohlii Birq. by meansof stem cuttings. The objectives were set, to study the effectof length, diameter and pretreatment by growth regulatoryhormones of stem cuttings, on its sprouting and survival.

MATERIALS AND METHODS

The experiment was conducted at the research farm ofT.C.B. College of Agriculture and Research Station, Bilaspur(Chhattisgarh), India during September 2008. Disease freeequally matured C. forskohlii plants were collected from themedicinal plant nursery established in this centre. Stemcuttings were prepared with two length sizes i.e. with twonodes, with four nodes and each type had three differentdiameter types viz. thick (0.65 cm), medium (0.52 cm), thin(0.43 cm).

Ganesh\Final\13

240

Three plant growth regulators i.e. Indole 3 acetic acid(IAA), -naphthalene acetic acid (NAA) and indole 3 butyricacid (IBA) were used in two (liquid and powder) formulations.As a liquid formulation IAA and NAA were applied at aconcentration of 500 ppm, 100 ppm, 50 ppm and 10 ppmwhereas, concentration of IBA was 1000 ppm, 500 ppm and100 ppm as per the usual recommendation [a]. Powderformulation was prepared at a concentration of 100 ppm and500 ppm for all the three growth regulators. For preparation ofpowder formulations, inert talc powder was mixedproportionately with the growth hormones to get the desiredconcentration. Water dipping (cuttings dipped in steriledistilled water) and control (without any treatment) treatmentswere added to compare the effect of growth regulators. ‘Dilutesolution dip method’ was used to apply the exogenoushormone formulations onto stem cuttings [9]. In brief, stemcuttings were dipped in the respective hormone preparationsfor overnight at room temperature and planted next day in thenursery bed. Irrigation water was applied for 24 hours byoverhead sprinklers during the experimental period. After 25days of planting the number of sprouted cuttings, number ofnode sprouted were counted whereas, their survival wasrecorded at 60 days after plantation. All the recorded datawere expressed in percentage.

The experimental design was randomized complete blockdesign with ten replications for each treatment. Data gatheredwere analyzed statistically using analysis of variance (ANOVA)and CD were calculated at 5% to separate the treatment means.

RESULTS AND DISCUSSION

Presented data indicated that the growth hormones atdifferent concentrations used as both formulations (liquid andpowder) along with the length and diameter of stem cuttingshad significant (Pd”0.05) effect on sprouting, nodes sproutingand per cent survival (Tables 1 and 2). Data presented in tableindicated that some of the concentrations of growth hormoneswere found to increase the sprouting significantly however,results were not consistent on the application of differentformulations. Moreover, irrespective of all treatments,exogenous hormone application induced mean sprouting,node sprouting and survival superiorly in powder formulationsas 44.7%, 22.9%, 83.7% than liquid formulations as 47.4%,24.4%, 89.4% (Tables 1 and 2).

Among growth hormones, IBA @ 100 ppm was foundsuperior (Pd”0.05) effectivity than the control, in enhancingthe growth as can be observed by the mean per cent sprouting,per cent node sprouting and per cent survival of 53.5%, 24.6%and 100.0% respectively in liquid formulation and 54.6%,26.3% and 100% respectively in powder formulation. Similarly,in powder formulation of IAA @ 500 ppm, the growth of cuttingwas superior to control (Figures 2 and 3). IBA were inconformity with the reports of its effectiveness as compared

to several naturally occurring auxins in promotion ofadventitious roots [9]. Moreover, Butola and Badola (2004)have recommended IAA and IBA as promising treatments toimprove rooting, growth and biomass in A. glauca andHeracleum candicans [3]. Hormone application mean in liquidformulation irrespective of other parameters revealed that IAA@ 100 ppm stimulated per cent sprouting (44.4%) and percent node sprouting (18.4%) comparable to water treatmentand control whereas, for IBA @ 500 ppm and 1000 ppm percent node sprouting (16.7%, 20.8%) and per cent survival(95.8%, 100.0%) was comparable with control (Figure 1). Incase of powder formulation application, per cent nodesprouting and per cent survival were comparable with controltreatment for NAA @ 1000 ppm (26.5%, 100.0%) NAA @ 500ppm (26.7%, 100.0%). Securedex B was least in enhancing thesprouting and survival of the stem cuttings compared to othertreatments (Figure 2). Chaudhry and Khan (2000) reportedthat IAA is known to promote the expansion of roots in cuttingthereby increasing the survival rate. Similarly, Choudhury andRashid (2000) reported IAA and IBA increase stem diameterwhereas, Sun et al. (1998) evaluated the effects of plant growthregulators (NAA, IAA, IBA) on sprouting of stem cuttings[5, 16].

0102030405060708090

100

IAA

500

ppm

IAA

100

ppm

IAA

50 p

pmIA

A 10

ppm

NAA

500

ppm

NAA

100

ppm

NAA

50 p

pmNA

A 10

ppm

IBA

1000

ppm

IBA

500

ppm

IBA

100

ppm

Wat

er d

ippi

ng

Con

trol

Treatments

Mea

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(%)

Figure 1. Bars represents mean per cent sprouting ( ), percent node sprouting ( ) and per cent survival ( )of stem cutting of Coleus forskohlii irrespective oflength, diameter of cutting after hormone applicationin liquid formulation

020

40

60

80100

120

IAA

1000

ppm

IAA

500

ppm

NAA

1000

ppm

NAA

500

ppm

IBA

1000

ppm

IBA

500

ppm

Wat

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Cont

rol

Treatments

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(%)

Fig. 2: Bars represents mean per cent sprouting ( ), percent node sprouting ( ) and per cent survival ( )of stem cutting of Coleus forskohlii irrespective oflength, diameter of cutting after hormone applicationin powder formulation.

241

Table 1. Effect of length of cuttings, diameter of cuttings and exogenous growth hormones as liquid formulations on per centsprouting, node sprouted and survival of stem cuttings of Coleus forskohlii

4 Node 2 Node Mean (2 Node & 4 Node)

ppm Dm Cutting

sprouted Node

sprouted Plant

survival Cutting sprouted

Node sprouted

Plant survival

Cutting sprouted

Node sprouted

Plant survival

(%) (%) (%) (%) (%) (%) (%) (%) (%) 500 D1 42.5 10.4 50.0 32.5 10.4 42.5 37.5 10.4 46.3

D2 33.3 8.3 50.0 50.0 25.0 50.0 41.7 16.7 50.0 D3 66.7 16.3 100.0 32.5 10.4 42.5 49.6 13.3 71.3

100 D1 33.3 8.3 42.5 33.3 12.5 47.5 33.3 10.4 45.0 D2 33.3 12.5 50.0 58.3 29.2 100.0 45.8 20.8 75.0 D3 50.0 18.8 50.0 58.3 29.2 100.0 54.2 24.0 75.0

50 D1 41.7 10.4 100.0 33.3 18.8 45.0 37.5 14.6 72.5 D2 58.3 14.6 100.0 33.3 12.5 47.5 45.8 13.5 73.8 D3 33.3 10.4 50.0 58.3 29.2 100.0 45.8 19.8 75.0

10 D1 41.7 10.4 100.0 16.7 10.4 100.0 29.2 10.4 100.0 D2 58.3 10.4 100.0 29.2 10.4 100.0 43.7 10.4 100.0

Indo

le A

cetic

aci

d

D3 22.9 10.4 100.0 58.3 20.8 100.0 40.6 15.6 100.0 500 D1 33.3 10.4 47.5 41.7 16.7 40.0 37.5 13.5 43.8

D2 25.0 14.6 40.0 25.0 14.6 45.0 25.0 14.6 42.5 D3 27.5 10.4 47.5 29.2 14.6 50.0 28.3 12.5 48.8

100 D1 58.3 14.6 100.0 29.2 10.4 50.0 43.7 12.5 75.0 D2 41.7 10.4 100.0 50.0 14.6 100.0 45.8 12.5 100.0 D3 25.0 8.3 100.0 29.2 10.4 50.0 27.1 9.4 75.0

50 D1 29.2 10.4 100.0 29.2 10.4 100.0 29.2 10.4 100.0 D2 66.7 16.3 100.0 29.2 12.5 100.0 47.9 14.4 100.0 D3 58.3 14.6 100.0 58.3 29.2 100.0 58.3 21.9 100.0

10 D1 25.0 10.4 100.0 16.7 8.3 100.0 20.8 9.4 100.0 D2 36.7 10.4 73.3 16.7 10.4 100.0 26.7 10.4 86.7

a-N

apht

hale

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cetic

aci

d

D3 50.0 12.5 25.0 50.0 25.0 50.0 50.0 18.8 37.5 1000 D1 58.3 29.2 100.0 58.3 29.2 100.0 58.3 29.2 100.0

D2 25.0 12.5 100.0 25.0 12.5 100.0 25.0 12.5 100.0 D3 41.7 20.8 100.0 41.7 20.8 100.0 41.7 20.8 100.0

500 D1 29.2 10.4 100.0 25.0 12.5 100.0 27.1 11.5 100.0 D2 25.0 10.4 100.0 25.0 10.4 100.0 25.0 10.4 100.0 D3 58.3 14.6 100.0 66.7 41.7 75.0 62.5 28.1 87.5

100 D1 41.7 10.4 100.0 41.7 10.4 100.0 41.7 10.4 100.0 D2 50.0 25.1 100.0 58.3 29.2 100.0 54.2 27.1 100.0

Indo

le B

uter

ic a

cid

D3 29.2 10.4 100.0 100.0 62.5 100.0 64.6 36.5 100.0 D1 58.3 20.0 100.0 41.7 20.0 100.0 50.0 20.0 100.0 D2 41.7 10.0 100.0 29.2 10.0 100.0 35.4 10.0 100.0 Water

Dipping D3 58.3 25.0 100.0 41.7 30.0 100.0 50.0 27.5 100.0 D1 58.3 15.0 100.0 0.0 0.0 0.0 29.2 7.5 50.0 D2 25.0 5.0 0.0 25.0 10.0 100.0 25.0 7.5 50.0 Control D3 83.3 20.0 100.0 66.7 40.0 100.0 75.0 30.0 100.0

Mean 41.3 16.1 81.5

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Table 3. CD at 5% level of different treatments for hormone application in liquid and powder formulations

C.D. at 5 % (liquid formulation)

C.D. at 5 % (powder formulation)

Treatments Sprouting Node sprouting

Survival Sprouting Node sprouting

Survival

Hormones 9.5 5.6 13.7 9.6 11.6 4.9

Diameter 4.1 2.7 7.4 9.4 4.6 5.1

Number of nodes 5.7 1.9 ns 3.5 3.8 2.6

Hormones x Diameter 16.9 9.5 15.7 12.9 13.8 9.5

Hormones x Nodes 13.8 7.7 12.8 10.5 11.3 7.7

Diameter x Nodes 5.8 5.4 5.4 6.1 6.5 4.5

Hormones x Diameter x Nodes 23.9 13.4 22.2 18.2 19.5 13.4

Table 2. Effect of length of cuttings, diameter of cuttings and exogenous growth hormones as powder formulations on per centsprouting, node sprouted and survival of stem cuttings of Coleus forskohlii

4 Node 2 Node Mean (2 Node & 4 Node) ppm Dm Cutting

sprouted Node

sprouted Plant

survival Cutting sprouted

Node sprouted

Plant survival

Cutting sprouted

Node sprouted

Plant survival

(%) (%) (%) (%) (%) (%) (%) (%) (%) 1000 D1 20.0 5.0 100.0 40.0 30.0 100.0 30.0 17.5 100.0

D2 80.0 32.5 100.0 37.5 20.0 87.5 58.8 26.3 93.8 D3 65.0 25.0 100.0 60.0 40.0 100.0 62.5 32.5 100.0

500 D1 50.0 15.0 100.0 20.0 10.0 100.0 35.0 12.5 100.0 D2 90.0 32.5 100.0 90.0 32.5 100.0 90.0 32.5 100.0

Indo

le A

cetic

aci

d

D3 90.0 35.0 100.0 80.0 40.0 100.0 85.0 37.5 100.0 1000 D1 30.0 11.5 100.0 32.5 13.5 100.0 31.3 12.5 100.0

D2 30.0 9.0 100.0 60.0 47.5 100.0 45.0 28.3 100.0 D3 90.0 32.5 100.0 60.0 45.0 100.0 75.0 38.8 100.0

500 D1 30.0 50.0 100.0 10.0 10.0 100.0 20.0 30.0 100.0 D2 50.0 22.5 100.0 20.0 10.0 100.0 35.0 16.3 100.0

a-N

apht

hale

ne a

cetic

ac

id

D3 70.0 22.5 100.0 60.0 45.0 100.0 65.0 33.8 100.0 1000 D1 22.5 10.0 100.0 22.5 10.0 100.0 22.5 10.0 100.0

D2 80.0 25.0 100.0 20.0 15.0 100.0 50.0 20.0 100.0 D3 60.0 22.5 100.0 50.0 45.0 100.0 55.0 33.8 100.0

500 D1 55.0 20.0 100.0 17.5 10.0 100.0 36.3 15.0 100.0 D2 80.0 27.5 100.0 30.0 15.0 100.0 55.0 21.3 100.0

Indo

le B

uter

ic a

cid

D3 100.0 40.0 100.0 45.0 45.0 100.0 72.5 42.5 100.0 D1 20.0 10.0 62.5 30.0 15.0 50.0 25.0 12.5 56.3 D2 40.0 15.0 52.5 30.0 25.0 50.0 35.0 20.0 51.3

Water Dipping

D3 50.0 22.5 100.0 70.0 35.0 60.0 60.0 28.8 80.0 D1 20.0 27.5 50.0 20.0 15.0 60.0 20.0 21.3 55.0 D2 30.0 12.5 67.5 35.0 15.0 20.0 32.5 13.8 43.8

Control

D3 42.5 27.5 75.6 42.5 30.0 57.5 42.5 28.8 66.6 Mean 47.4 24.4 89.4

243

36.5 37.5

49.8

13.1 13.921.4

79.4 82.0 82.6

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100

Thick Medium Thin

Branch cutting diameter (cm)

Mea

n va

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(%)

Fig. 3: Bars represents mean per cent sprouting ( ), percent node sprouting ( ) and per cent survival ( )of stem cutting of Coleus forskohlii at three differentcutting diameter (thick= 0.65 cm, medium= 0.52 cm,thin= 0.43 cm) irrespective of cutting length afterhormone application in liquid formulation.

27.5

50.2

64.7

16.422.3

34.5

88.9 86.193.3

0102030405060708090

100

Thick Medium Thin

Brach cutting diameter (cm)

Mea

n va

lues

(%)

Fig. 4: Bars represents mean per cent sprouting ( ), percent node sprouting ( ) and per cent survival ( )of stem cutting of Coleus forskohlii at three differentcutting diameter (thick= 0.65 cm, medium= 0.52 cm,thin= 0.43 cm) irrespective of cutting length afterhormone application in powder formulation.

Mean values of per cent sprouting, per cent nodesprouting and per cent survival irrespective of hormone type,concentration and length of cutting revealed that the thinstem cuttings were significantly more inducible by theapplication of hormones in both liquid and powderformulations whereas, growth of thick cuttings were leastinducible (Figures 3 and 4). For example, mean per centsprouting was significantly higher in thin cuttings (49.8%)than thick (36.5%) after liquid formulation application. Similarly,after application of liquid formulation sprouting mean wasconsiderably higher in thin cuttings (64.7%) than thick cuttings(27.5%). Same trends were followed for mean per cent nodesprouting and mean percent survival of stem cuttings (Figures3 and 4). Exogenous hormone applications in powerformulations were more effective than liquid formulations onaverage. For example, the average per cent sprouting, nodesprouting and survival of thin stem cuttings in powder

formulations 64.7%, 34.5%, 93.3% were noticeably higher thanliquid formulations 49.8%, 21.4%, 82.6% respectively (Fig. 3& 4).

Coleus forskohlii is propagated by stem tip cuttings,described as herbaceous cuttings [8]. Market demand of thesespecies for pharmaceuticals and ethno-medicinal utility, aremet through harvesting from wild populations. Thus, plantgrowth regulators can be widely used in vegetativepropagation to improve sprouting, rooting and survival ofcuttings. From the present study it can be concluded thatIndole 3 Acetic Acid (IAA), - naphthalene acetic acid (NAA)in powder formulation are applicable to thin stem cuttings ofColeus forskohlii for better sprouting and absolute survival.

ACKNOWLEDGEMENT

The authors are thankful to Chhattisgarh Council ofScience and Technology (CGCOST) for providing the requisitefunds to carry out the research work which is part of theproject entitled ‘Conservation of indigenous medicinal plantof Chhattisgarh forest with special reference to growingbehavior of commercially important species’.

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