9ECCD7E7d01

EFFECT OF PLANT GROWTH REGULATORS ON GROWTH AND

YIELD OF PATCHOULI (Pogostemon cablin Benth. L.)

Thesis submitted to the

University of Agricultural Sciences, Dharwad

in partial fulfillment of the requirements for the

Degree of

Master of Science (Agriculture)

in

CROP PHYSIOLOGY

By

ANILKUMAR M.

DEPARTMENT OF CROP PHYSIOLOGY COLLEGE OF AGRICULTURE, DHARWAD

UNIVERSITY OF AGRICULTURAL SCIENCES,

DHARWAD - 580 005

AUGUST, 2005

ADVISORY COMMITTEE

DHARWAD (A.S. NALINI PRABHAKAR) AUGUST, 2005 MAJOR ADVISOR

Approved by :

Chairman : ____________________________

(A. S. NALINI PRABHAKAR)

Members : 1. __________________________

(R. V. KOTI)

2. __________________________

(S. M. HIREMATH)

3. __________________________

(A. A. PATIL)

C O N T E N T S

Chapter No.

Title Page No.

I INTRODUCTION

II REVIEW OF LITERATURE

III MATERIAL AND METHODS

IV EXPERIMENTAL RESULTS

V DISCUSSION

VI SUMMARY

VII REFERENCES

LIST OF TABLES

Table No.

Title Page No.

1. Monthly meteorological data for the experimental year (kharif, 2004) and the mean of past 54 years (1950-2003) as recorded at the Meteorological Observatory, Main Agricultural Research Station, University of Agricultural Sciences, Dharwad (Karnataka)

2. Physical and chemical properties of the soil of experimental area

3. Salient feature of the growth regulators and chemicals used in experiment

4. Influence of plant growth regulators on plant height (cm) in patchouli

5. Influence of plant growth regulators on number of branches in patchouli

6. Influence of plant growth regulators on number of leaves in patchouli

7. Influence of plant growth regulators on leaf dry weight (g/plant) in patchouli

8. Influence of plant growth regulators on stem dry weight (g/plant) in patchouli

9. Influence of plant growth regulators on total dry matter (g/plant) in patchouli

10. Influence of plant growth regulators on leaf area (dm²/plant) in patchouli

11. Influence of plant growth regulators on leaf area index (LAI) in patchouli

12. Influence of plant growth regulators on leaf area duration (days) in patchouli

Contd…..

Table No.

Title Page No.

13. Influence of plant growth regulators on leaf area ratio (cm²/g/plant) in patchouli

14. Influence of plant growth regulators on specific leaf area (cm²/g) in patchouli

15. Influence of plant growth regulators on specific leaf weight (mg/cm²) in patchouli

16. Influence of plant growth regulators on absolute growth rate (AGR, g/day) in patchouli

17. Influence of plant growth regulators on crop growth rate (CGR, g/dm²/day) in patchouli

18. Influence of plant growth regulators on relative growth rate (RGR, g/g/day) in patchouli

19. Influence of plant growth regulators on net assimilation rate (NAR, mg/cm²/day) in patchouli

20. Influence of plant growth regulators on number of oil glands (glands/mm² leaf area) in patchouli

21. Influence of plant growth regulators on chlorophyll ‘a’ and ‘b’ content (mg/g fresh weight) in patchouli

22. Influence of plant growth regulators on total chlorophyll (mg/g fresh weight) in patchouli

23. Influence of plant growth regulators on carotenoid content (mg/100 g fresh weight in patchouli

24. Influence of plant growth regulators on phenols content (mg per g dry weight) in patchouli

25. Influence of plant growth regulators on nitrate reductase

activity NRA (µ moles of NO2 fixed/g fresh weight) in patchouli

26. Influence of plant growth regulators on yield parameters of patchouli (FWB and DWB)

LIST OF FIGURES

Figure No.

Title Between pages

1. Plan of layout

LIST OF PLATES

Plate No.

Title Between pages

1. General view of the experimental site

I. INTRODUCTION

Aromatic plants are the plants which contain essential oils. These essential oils or volatile oils as they are often called, are found in different species of plants which are known to be very complex in their chemical nature. These are the mixture of acyclic and cyclic monoterpenoids, the physiological significance of which as far as the plant is concerned is not obvious. These probably represent by products of metabolism rather than foods and they have no apparent function in the plants with regard to primary metabolism.

The characteristic flavour and aroma that they impart are basically advantageous in

attracting insects and other animals which play a role in pollination or dispersal of seeds and fruits. However, the aroma in these essential oils is being exploited largely in perfumery, cosmetic and pharmaceutical industries.

India has a perfumery tradition that dates back to over 5000 years to Indus valley

civilization. In the excavations of Harappa and Mohanjodaro, a “water distillation still” and “receiver” have been recorded whose shape resemble to the “deg” and “bhaka” currently used by “attars” (traditional perfumers) of kannauj in India.

Bucchbauer (1990) has recorded examples of several aromatic plants presently in

use for medication which have come to us from our ancestors inhabiting different countries. Thus, ‘anise oil’, ‘citronella oil’, ‘eucalyptus oil’, ‘spruce oil’ and aroma chemicals like camphor, menthol, cineol, thymol and guacacol are still used as both aromatic and additive. In addition to the aromatic oils, the finer perfumes contain fixatives substances, which are less volatile than the oils and delay equalize evaporation. There are several aromatic species which are utilized for this purpose. Patchouli is one among them.

The patchouli plant was first described by Pelletier Scutelet and named Pogostemon

patchouli. In 1986, Holmes identified it as Pogostemon cablin Benth. a native of the Philippine islands, the word “Cablin” being arrived from “Cablam”, vernacular name of the plant in the Philippines. The plant has also been described by Blanco in his Flores de Filipinos as Mentha cablin.

Today the true patchouli plant of commerce, Pogostemon cablin Benth. (syn. P

Patchouli pellet) belonging to family Lamiaceae, serves for distilling the essential oil is cultivated in British Malaya (Straits Settlements and Johare State) and more extensively in Indonesia (Northern Sumatra). The Malayan Vernacular term “Dhulam wangi or tilam wangi” means sweet or cultivated patchouli.

Patchouli is grown mainly in Indonesia, Malaysia, China and to lesser extent in

Madagascar, Reunion and Seychelles. Though efforts have been made by several institutions since early fifties, current indigenous production of patchouli oil is hardly few hundred kilograms and that too quality variation is very large. Indonesia is producing about 600 tonnes and China about 30 to 40 tonnes of oil. Indian consumption is about 40 tonnes of pure oil and about 70 tonnes of formulated oil. International prices have recently gone up to US $20 to 25 per kg, therefore landed cost would be higher than Rs.1500 per kg. Thus, there is a large potential for patchouli oil production in India.

The commercial oil of patchouli is obtained by steam distillation of the shade dried

leaves and is one of the most important naturally occurring essential oil used in perfumery industry. Although rarely used as dominant source of fragrance in its own right, the oil is widely used to give a solid foundation and lasting character to a fragrance. Patchouli oil has notably strong fixative properties and helps to prevent rapid evaporation of a perfume and thereby promotes, tenacity and its dominant woody note, although the aroma possesses other characteristics and is very complex. The oil is generally blended with other essential oils, such as geranium or clove before use.

Patchouli oil is used in a wide range of toilet soaps, scents, lotions, pre-shave and

after-shave lotions and detergents. Its strong tenacity render it to be particularly suitable for

heavy perfumes and for imparting a lasting character and strength to lighter perfumes. In very low concentration (0.002% or 2.21 ppm) the oil is extensively used as a flavour ingredient in major food products including alcoholic and non alcoholic beverages, frozen dairy desserts, candy, baked goods, gelatin, meat and meat products. Blended with sandalwood, it gives one of the finest attars, widely used in soaps, cosmetics, tobacco and incense sticks.

Dry patchouli leaves are used for scenting wardrobes. The leaves and tops are added

in bath for their antirheumatic action. In Chinese medicine decoction from the leaves are used with other drugs to treat nausea, vomiting, diarrhoea, cold and headaches (Leiung, 1980). A related species patchouli (Pogostemon heyneanus) is reported to content principle possessing anticancer activity (Purushothaman et al., 1985).

Pogostemon cablin yields patchouli oil, which has a great demand in the perfume and

flavour industry. At present, the global requirement of patchouli is met mainly through production from Indonesia. However, due to adverse conditions in Indonesia, the supply of oil is irregular. India’s available infrastructure and environment can provide an opportunity to gain a major part of the world market. The global trade in patchouli oil is to the tune of 200 crores (1000 tonnes) per year. Most of which presently comes from Indonesia. The Indian perfumery market consumes 70-80 tonnes per annum and there is good potential for export, which focussed programmes for promoting cultivation of this plant, thus India can become the second largest producer of patchouli oil in the world.

Commercial cultivation of the crop in India was first attempted by Tata Oil Mills in

1942 (Kumar et al., 1986). After initial stray attempt to grow the crop, its systematic cultivation started in 1962 by CIMAP (Kumar et al., 1986). Patchouli can be cultivated in coastal regions of India as a main crop or as an intercrop along with plantation crops. India which has started cultivating patchouli is importing annually about 20 tonnes of pure patchouli oil and 100 tonnes of formulated oil which is certainly a very meagre quantity. This appears to be mainly because of non availability of planting material to the farmers. Inspite of farmers enthusiasm to cultivate patchouli, inadequate supply of planting material is one of the major setback. Similarly, farmers inability to increase the yield potentiality down the production of patchouli oil has ultimately brought down the production of patchouli oil.

The plant growth regulators (PGR’s) are considered as a new generation

agrochemicals after fertilizers, pesticides and herbicides. Plant growth regulators are the chemical substances, when applied in small amounts

modify the growth of plants usually by stimulating or inhibiting part of the natural growth regulatory system. About sixty plant growth regulators are now commercially available and several of them have reached considerable importance in crop production. The growth regulators include both growth promoters and retardants which have been shown to modify the canopy structure and other yield attributes.

Plant growth regulators are known to have great potential ability to increase the

productivity of horticulture crops. The response of plant or plant parts to growth regulators varies due to fluctuations in endogenous hormonal level of the plant and the manner in which the natural growth regulators interact with the applied growth regulator.

Though the PGR’s have great potentialities to influence plant growth and

morphogenesis, its application and actual assessments etc. have to be judicially planned in terms of optimal concentrations, stage of application, species specificity and seasons etc. These constitute major impediments of PGR’s applicability. In view of this an investigation was conducted to study the effect of plant growth regulators on growth and yield of patchouli with following objectives.

1. To study the effect of different concentrations of growth regulators on morphological

and biochemical characteristics of patchouli 2. To assess the influence of growth regulators on growth and anatomical

characteristics 3. To study the effect of growth regulators on yield and yield components of patchouli

II. REVIEW OF LITERATURE

Patchouli is known to be one of the popular aromatic crops with immense commercial importance. Inspite of its economic potentiality much work has not been done using growth regulators to increase productivity. Therefore, the present investigation was undertaken in order to test the effect of plant growth regulators on growth and yield of patchouli.

In patchouli, most of the work has been done on the influence of PGRs on

propagational aspects, the studies with reference to influence of PGRs on growth and yield, however is very much limited. The available information on effect of different plant growth regulators on growth and yield of patchouli as well as other aromatic plants have been reviewed and presented.

2.1 Morphological parameters (plant height, number of branches, number of leaves)

Misra (1995) reported that application of 10-500 mg GA3/litre to shade grown

patchouli cv. Johor plants increased the number of nodes, branches, number of green leaves, total leaf area, specific leaf fresh weight, leaf area index, photosynthetic pigment (chl a and b and carotenoid) contents and essential oil yield but had no effect on plant height, leaf fresh weight and number of yellow leaves. The optimum concentration of GA3 for least growth pigment content and oil yield was with 250 mg/litre.

Mahmoud et al. (1996) studied the response of growth and essential oil content of

sweet basil (Ocimum barilicum L.) to some natural hormones GA3, IAA and kinetin. According to them GA3 stimulated plant growth (plant height, number of leaves, leaf area, and FW and DW) compared to other growth regulators. However, decreased oil content was noticed in GA3 while IAA and kinetin increased oil content.

According to Nilsmranchit et al. (1996) GA3 treatments, 1 or 2 months before the

harvest increased the number of leaves, but slightly decreased tanin concentrations in case of Geranium thunbergii Sieb.

Bhat et al. (1989) conducted investigation to test the effects of GA3. CCC and TIBA

on the growth, herbage yield and essential oil content in Davana (Atremesia pallens Wall) which revealed that plants treated with GA3 showed increase in plant height, number of branches, plant spread as well as fresh and dry weight of flower heads and herbage. The essential oil yield was significantly enhanced by GA3 at 100 ppm and less so by TIBA and CCC at 400 and 4000 ppm respectively. Further, early flowering was observed with GA3 treatment, while it was found to be delayed when CCC or TIBA were applied. These inhibitors also suppressed plant height significantly.

Vasundhara et al. (1992) conducted studies to find out the influence of GA3, cycocel

and Triacontanol on growth, herbage, oil content and oil yield in Marjoram. They revealed that plants treated with GA3 showed an increase in plant height, number of branches and plant spread, while cycocel and Triacontanol at higher concentrations held a negative effects on all these parameters. Application of GA3 at 2000 ppm was found to increase the fresh herb recovery recording the maximum oil yield. Although cycocel at 50 ppm and Triacontanol at 6 ppm increased the fresh herb recovery and oil yield over control but were next to GA3.

A four years study conducted by Balyan et al. (1994) on lemon grass (ckp-25)

showed that application of Triacontanol at lower concentrations upto 10 ppm significantly increased the plant height, number of leaves and leaf length. It had positive effect on herbage yield which increased by 21.35 per cent over control. Higher concentration of 60 and 80 ppm had negative effect on herbage yield and oil content, hence did not get influenced by Triacontanol application.

Studies conducted by Bhattacharya and Rao (1996) on effect of Triacontanol and mixatalol on rose scented geranium (Pelargonium sp.) revealed that foliar application of long chain aliphatic alcohol plant growth regulators such as triacontanol and mixatalol significantly increased yield attributes of Pelargonium cv. Bourbon including height, number of branches, weight of leaves and stems, branches, shoot and roots and essential oil composition yield and recovery. The enhancement over control plants in root yield was much greater than shoot yield indicating the influence of these plant growth regulators on root growth and development of leaf cuttings of third, fourth and fifth leaves producing adventitious roots and shoots under ambient conditions when treated (foliar spray) with Triacontanol. It also increased the number of root, leaf, petiole and shoot weights. Distillation of leaves after rooting, however yielded no essential oil.

Mohandas and Sampath (1985) made studies on the regulation of growth and yield in

geranium with some hormonal sprays, wherein height of plant was much influenced by GA3

and alar sprays. Significantly higher plant height was noted under GA3 250 ppm as compared to control. At higher concentrations of ethanol and CCC (400 ppm each) shortened it as against control. Significant increase in total plant spread was noticed with the spray of alar at 1000 ppm recording 12166.32 sq cm as against control registering only 6101.78 sq cm. The foliage yield in general increased with CCC @ 1000 ppm and alar @ 1000 ppm concentration sprays but ethrel spray at 4000 ppm caused significant reduction over control.

Umesha et al. (1991) studied influence of GA3 and cycocel (CCC) on growth and

productivity of clocium (Ocimum gratissimum L.) and revealed that while GA3 promoted plant height, internodal length, leaf area and dry matter accumulation, CCC had a negative influence on all these parameters. Although CCC application resulted in early flowering, leaf yields were considerably reduced. GA3 was found to increase yield of essential oil, eugenol content and eugenol yield.

Mohamed et al. (1983) conducted pot experiment to study effects of GA3 and

chloromequat on yield and oil quality of geranium in two successive seasons where in GA3 increased plant height and herb production. Oil percentage and oil yield, however decreased. On contrary chloromequat although decreased plant height, it increased both percentage and yield of oil.

Studies made by Gupta et al. (1992) on effect of growth regulators gibberlic acid and

tria contanol on supplying stage of Ocimum carnosum under field conditions indicated that Triacontanol although had promotory effect on plant height, leaf number and leaf area, gibberlic acid, however did not show any effect on growth parameter except plant height and internodal length. Total chlorophyll and protein contents were increased in leaf under the influence of Triacontanol, but gibberlic acid has no effect.

Krishnamoorthy and Madalagere (2000) noticed application of GA3 300 ppm in

ajowan (Trachyspermum ammi L.) increased plant height and recorded maximum number of secondary and tertiary branches followed by that of CCC and NAA. Maximum number of leaves, total dry matter and higher seed yield was recorded by GA3 300 ppm followed by GA3 200 and 100 ppm. The essential oil content of seeds increased with NAA application.

An experiment conducted by Singh and Bajimol (2001) to study effect of plant

bioregulators GA3, ethrel and kinetin at different concentrations on growth flowering and bulb production in tuberose has revealed the maximum number of sprouts per clump, length of spike and duration of flowering with 100 ppm GA3 where as 200 ppm GA3 significantly increased number of leaves/plant, plant height, number of florets/spike and weight of florets/plant. In general, all the PGRs has significantly influenced on various parameters over control except any conspicuous effect on bulb diameter and number of spikes/plant.

Gowda and Krishnan (1992) studied effect of three concentrations each of GA3, CCC

and kinetin on morphological and solasodine content of Solanum viarum Dunal. The results revealed that GA3 treatments increased plant height and internodal length at 60 days. Plant spread was increased at lowest concentration of GA3 @ 250 ppm.

In an experiment conducted by Borse and Dhumal (2001) to enhance solasodine contents in Solanum kharianum, GA, IAA and CCC were used as foliar sprays in different concentrations. Results revealed that all treatments increased plant height, number of branches and leaves per plant and reduced spines on leaves and stem.

Shedeed et al. (1990) reported increased leaf number, leaf area, FW and DW in case

of basil plant by GA3 and kinetin application and optimum concentration of GA3 was 100 ppm and for kinetin it was 50 ppm.

Shukla et al. (1992) observed increased plant height, artemirinin level and shoot fresh

weight in case of Artemisia annua L. plants treated with triacontanol at 1.0 and 1.5 mg/litre. According to Srivastava and Sharma (1991) the plant height, capsule number and

weight, morphine content, CO2 exchange rate, total chlorophyll and shoot fresh and dry weight were greatest at triacontanol (0.01 mg/ltr) treated opium poppy (Papaver somniferum L.) plants.

According to Shahine et al. (1992) in the plants of fenugreek (Trigonella foenum

graceum) sprayed with growth regulators, gibberlic acid increased leaf area, leaf number and stem length. Paclobutrazol and ethrel increased branching and leaf chlorophyll, but carotenoid concentrations were decreased by gibberlic acid and ethrel.

2.2 Anatomical parameters

Singh and Hippalgaonkar (1993) studied the effect of three concentration of kinetin on the growth characteristics and oil yield of patchouli. It was revealed that, kinetin treatment showed an increase in axillary bud development, number of leaves, total leaf area/plant, herbag yield, chlorophyll content, leaf gland number and oil yield. Of the three kinetin concentrations tested 0.5 x 10

-4 M kinetin was found most effective in improving growth and

increasing oil yield of patchouli. According to Pappaiah and Muthuswamy (1980) five growth regulators viz., GA3 (25

ppm), MH (1000 ppm), TIBA (100 ppm), cycocel (800 ppm) and ethrel (100 ppm) on three year old Jasminum grandiflorum bushes to study the histological changes in the leaf. GA3 and ethrel reduced the leaf thickness and length of palisade layers and the width of epidermal layer was increased by MH and TIBA appreciably.

2.3 Physiological and biochemical parameters (chlorophyll, nra, cartenoid content)

Kanjilal and Singh (1998) studied the effect of four concentrations (25, 50, 100 and

150 mg/l) of gibberlic acid (GA3) indole-3-acetic acid (IAA) and 1-napthalene acetic acid (NAA) with a control on flower heads essential oil, total chlorophyll, carotenoids and protein content of chamomite (Chamomilla recutita L.). They observed that increasing the concentration of IAA and NAA resulted a progressive reduction of plant height, yield of dry flower heads and essential oil, chlorophyll at 50, 100 and 150 mg L

-1 of IAA and control

respectively. Content of chlorophyll increased in the GA3 treated plants over the lower concentration. On the other hand, the content of chlorophyll remained low in increasing doses of IAA and NAA. The fairly high content of protein (42.5 g kg

-1) was observed in 100 mg L

-1 of

IAA. The protein content decreased with increased NAA concentration. Studies made by Singh and Misra (2001) on effect of GA3 and ethrel at high

concentration (1000 microgram/m) on Mentha spicata var. M35-5 observed enhancement in fresh weight biomass, leaf stem ratio, specific leaf weight and chlorophyll content. Nitrate reductase activity and peroxidase activity with low oil content. Limonene increased significantly in ethrel and GA3 treated genotypes.

Application of 0.4 µg/ml increased micronutrient uptake, total chlorophyll and citral content in case of lemon grass (Cymbopogon flexosus), but chlorophyll a: b ratio and stomatal resistance was decreased (Misra and Srivastava, 1991).

2.4 Yield and yield parameters (total herbage yield, leaf yield, seed yield and essential oil yield)

A field trial was conducted by Bhaskar et al. (1997) to study the influence of growth

regulators such as TIBA and kinetin in 3 concentration viz., 25, 50 and 100 ppm and 50, 100 and 200 ppm respectively on the production and regeneration capacity of patchouli (Pogostemon patchouli) cultivar IIHR PP-1. Results indicated that maximum fresh herbage production of 3.23 t/ha and oil yield of 24.44 kg/ha in the first harvest was recorded with the application of TIBA at 100 ppm and kinetin at 200 ppm. Though maximum oil content of 2.53 per cent was observed with this treatment the differences were not significant. Application of kinetin alone at higher concentration of 200 ppm resulted in improvement in both herbage and oil yield. Both herbage production and oil content showed increasing trend with increase in concentration of either each regulator alone or in combination. However, it was also observed that regeneration was not improved with any of growth regulators or in combination.

Jadhav et al. (2003) conducted a pot culture experiment showing that stem cuttings of

10-15 cm length with 2-4 leaves and three internodes on dipping in IBA 1500 ppm for 30 seconds produced maximum number of leaves, length of shoot, leaf area, biomass of plant, while 30 seconds dipping in IBA 2000 ppm showed maximum oil content in patchouli (Pogostemon cablin Benth).

Tasma and Moko (1988) observed that triacontanol (0.5%) auxin (0.25) + cytokinin or

phenol compound (2 ppm) at two weekly intervals or 2,4-D (0.5%) at 4 weekly interval were found to enhance plant growth and yield with auxin cytokinin mixture having maximum effect in patchouli.

Singh et al. (1989) studied effect of growth regulator on rooting and yield of geranium.

They observed that maximum essential oil yield was obtained at IBA (500 ppm), further IBA (200 ppm) was proved to be optimum for oil characters and is recommended for better growth and yield.

Barua and Hazarika (1982) reported that the application of NAA in different

concentrations significantly affected leaves and solasodine content of Solanum khasrianum Clarke. Further they observed that NAA application at 50, 100, 150 and 200 ppm led to dark green leaves in horizontal and droopy portions and increase in solasodine content was observed in 50, 100 and 150 ppm concentration.

In the pot culture experiments conducted by Bhattacharya et al. (1995) in Geranium

to study the effect of foliar application of indole-3-acetic acid (100 mg/l), indole-3-butyric acid (100 mg/l) and cycocel (2000 mg/l) increased number and yield of leaves was exhibited. Shoot (leaves + stem) essential oil and concentration of essential oil of rose scented geranium cv. Bourbon was also noticed compared to control. Napthyl acetic acid (50 mg/l) and gibberlic acid (200 mg/l) did not influence the yield of rose scented geranium at the concentration tried with kinetin (100 mg/l). However, increased leaf and shoot yield was noticed but not essential oil yield. Stem yield (without leaves) of the crop was not affected by any of plant growth regulators.

A field experiment was conducted by Singh (2003a) to evaluate the response of plant

bio-regulators on growth and flowering in marigold. In his experiment plants of marigold were sprayed with GA3 (100 and 200 ppm), IAA (50 and 100 ppm), kinetin (50 and 100 ppm) and control with distilled water. Maximum fresh and dry leaf biomass were recorded with GA3 100 ppm treatment. Kinetin gave pronounced effect in increasing number of branches/plant and leaf area which were statistically on par with GA3 100 ppm. Earliness on flower bud initiation and flowering were noticed with IAA 50 ppm, followed by GA3 100 ppm. Lower value leaf biomass and flower yield were encountered with control.

Hegde et al. (2001) studied the effect of three phytoharmones, BA, NAA and GA3 at different concentrations (25 and 50 ppm) on Kasturbhendi. The results indicated that NAA at 25 and 50 ppm concentrations increased number of fruits/plant and total biological yield.

Figueiredo et al. (2002) evaluated the effect of plant growth regulators GA3 (50 and

100 mg/litre), ethrel (100 and 200 mg/litre); CCC (500 and 1000 mg/litre); alar 85 (1000 and 2000 mg/litre), accel (20 and 40 mg/litre) and control on citral yield of lemon grass. But, no plant growth regulator used increased citral content.

Sahhar et al. (1984) studied the effect of gibberlic acid on some botanical and

chemical characteristics of basil (Ocimum basilicum L.) where in herbage fresh weight and essential oil yield in all cases increased with GA3 at 90 ppm.

Mousa and Emary (1983) examined the effect of foliar applied gibberlic acid and

maleic hydrazide on sweet basil in which GA3 considerably reduced plant height but increased herbage yield. MH at 100 ppm greatly increased number of branches per plant while oil percentage of fresh herbage was significantly increased by GA3 at 50 ppm and MH at 100 or 200 ppm.

Kewalanand et al. (1998) studied effect of plant growth regulators on Japanese mint

(Mentha arvensis) var MAS-1 in which growth parameters, essential oil, content, fresh herbage and oil yield were noticed to be significantly enhanced by application of GA3 at 40 mg/litre in comparison with NAA and control.

A low dose of plant growth stimulant triacontanol (0.1 g/ml) to Mentha arvenis L.

increased essential oil yield as well as fresh and dry matter production, net photosynthetic rate (pn) chlorophyll content and chl a–b ratio (Srivastava and Sharma, 1991).

Ansari et al. (1988) recorded that IAA and GA3 applications in Cymbopogon

jawaraneusa increased leaf length, width and dry weight whereas treatment with IBA decreased all three values IAA and GA3 increased the essential oil yield as well as the content of major constituent piperitone, compared to IBA.

Randhawa et al. (1988) observed that sucker treatment with GA3, resulted in early

emergence and the production of higher number of sprouts ultimately giving higher herb and oil yields in Mentha arvensis.

Yaseen and Tajuddin (1998) recorded highest oil in case of Artemesia by application

of GA3 and IAA at 150 ppm. Further, Artemirin content was highest in 6 ppm triacontanol treated plants.

Gupta et al. (1995) analysed physiology of growth in Ocimum carnosum L. Kotto

treated with triacontanol wherein results revealed that TRIA 10 µg/ml increased leaf character, NAR, CGR and consequently increasing total biomass and herbage yield/plant.

According to Kadam et al. (1998) the foliar spray of 5.0 ppm vipul increased nitrate

reductase activity, essential amino acid in spinach leaves. Farooqui et al. (1996) noticed that application of GA3 (25 or 50 mg/litre) significantly

increased fresh weight yield, artemisin content and yield, and essential oil content (µl/g) in comparison with control plant in case of Artemisia annua L.

In a field trials conducted by Saeid et al. (1994) on effect of growth regulators on

herb, oil yield and hormonal content of lemon grass (Cymbopogon citratus) indicated that application of mepiquatchloride (125-500 ppm) and IBA (25-100 ppm) reduced the herb yield and oil yields/plant but percentage of oil contents were slightly increased compared to control.

Singh (2003b) reported that GA3 significantly increased fresh weight of leaf and

diameter of flower, however maximum fresh and dry weight of leaf biomass, leaf area index,

number and weight of flowers per plant were highest in case of GA3 200 ppm in calendula (Calendula officinalis).

Analysis of growth and development and essential oil formation in O.

kelimandscharicum revealed that GA3 augmented the essential oil biogenesis and the content and increased oil extension growth, herbage yield and also oil content in all concentration under experimental studies (Anonymous, 1998).

Rao et al. (1994) concluded that foliar application of plant growth regulators such as

ethephon (0.062%), cytozyme (0.3%), biozyme (0.1%) and chamatkar (0.3%) in geranium cv. Bourbon significantly increased the growth (plant height, number of branches/plant), biomass production (yield of leaves, stem and branches with cut leaves, shoot, root yield) and chemical profile of essential oil.

Sudria et al. (1999) mentioned that plantlets treated with plant growth regulators

showed increased oil accumulation in case of Lavandula dentata. However, growth of plantlets were not correlated with their essential oil content.

III. MATERIAL AND METHODS

A field experiment was conducted during kharif 2004 to study the effect of plant growth regulators on growth and yield of patchouli. The details of materials used and the experimental technique adopted during the course of investigation are described below.

3.1 Experimental site

The field experiment was conducted at Main Agricultural Research Station, University of Agricultural Sciences, Dharwad which is situated at 15°12’ N latitude 75°7’ E longitude and at an altitude of 678 m above the mean sea level. The experiment was laid out in plot number 125 of E block on medium black soils.

3.2 Climate

The main research station of University of Agricultural Sciences, Dharwad is situated in Northern Transitional Tract (Zone-8) of Karnataka state. The meteorological data of previous 54 years and the year of investigation recorded at the meteorological observatory, Main Agricultural Research Station, Dharwad are presented in Table 1.

3.3 Soil characteristics

The experimental site consisted of black clay loam soil. Composite soil samples were collected from the experimental site and analysed for various physiological and chemical properties and presented in Table 2.

3.4 Previous crop

The previous crop was watermelon.

3.5 Experimental details 3.5.1 Design and layout

The experiment was laid out in randomised block design with ten treatments in three replications. The plan of layout of the experiment is given in Figure 1.

3.5.2 Plot size

Gross plot size : 3 m x 3 m² Net plot size : 1.8 m x 1.8 m²

3.5.3 Description: Patchouli (Pogostemon cablin Benth.) cultivar genotype Johore

The cultivar Johore used in the experiment is a perennial, profusely branched, erect

or ascending aromatic herb/under shrub, pubescent with quadrangular stem, leaves are simple, opposite, decussate pale to purplish green when grown in open field condition. Its odour is woody earthy followed by aromatic spicy.

Table 1. Monthly meteorological data for the experimental year (kharif, 2004) and the mean of past 54 years (1950-2003) as recorded at the Meteorological Observatory, Main Agricultural Research Station, University of Agricultural Sciences, Dharwad (Karnataka)

Temperature (°C) Rainfall (mm)

Mean maximum Mean minimum Relative humidity (%)

Month

2004 Mean* 2004 Mean* 2004 Mean* 2004 Mean*

January - 0.086 29.6 29.15 14.7 19.23 54 63.34

February - 1.161 32.5 34.52 16.4 16.02 53 51.18

March - 0.147 36.5 35.73 19.6 18.81 49 56.47

April 24.4 48.45 37.4 37.00 19.8 21.32 51 76.98

May 61.1 81.40 33.6 36.52 21.4 21.48 66 66.71

June 43.8 109.14 28.8 29.50 21.5 21.21 80 81.69

July 24.8 145.77 29.2 22.06 21.0 20.95 79 87.46

August 160.7 95.30 27.0 22.01 20.3 20.62 83 86.51

September 222.1 100.54 28.6 28.75 19.9 20.16 77 82.40

October 64.6 130.99 30.1 30.12 18.4 19.30 65 76.44

November 0.6 32.04 30.2 29.46 15.9 15.50 52 68.13

December 0.0 54.50 29.4 29.18 12.5 13.44 45 63.81

Total 602.1 750.43

Mean of 54 years (1950-2003)

Table 2. Physical and chemical properties of the soil of experimental area

Sl. No.

Particulars Values Rating Method employed

A. Physical properties

1. Mechanical analysis

Coarse sand (%) 6.28 Clay International pipette method (Piper, 1966)

Fine sand (%) 14.27 Clay International pipette method (Piper, 1966)

Silt (%) 27.52 Clay International pipette method (Piper, 1966)

Clay (%) 51.99 Clay International pipette method (Piper, 1966)

2. Bulk density (Mg m-3) 1.33 Core Sampler Method (Dastane, 1967)

B. Chemical properties

1. Soil pH (1:2.5 soil water extract)

7.6 Slightly alkaline

pH meter (Piper, 1966)

2. Electrical conductivity (dSm-1)

0.28 Normal Conductivity Bridge (Jackson, 1973)

3. Organic carbon (%) 0.52 Medium Wet oxidation method (Jackson, 1967)

4. Available nitrogen (N) (kg ha-1)

221 Low Alkaline permanganate method (Subbaiah and Asija, 1959)

5. Available phosphorus (P2O5) (kg ha-1)

32.4 Medium Olsen’s method (Jackson, 1967)

6. Available potassium (K2O) (kg ha-1)

318.7 High NH4OAC Extract method (Jackson, 1973)

Table 3. Salient feature of the growth regulators and chemicals used in experiment

Sl. No.

Common name Chemical Chemical name Physiological effect

1. NAA Awan NAA Growth promoter, stimulates cell division, cell elongation and elongation of shoot, photosynthesis, RNA synthesis, membrane permeability to water uptake, prevents abscission of to leaves, enhance leaf area index leaf chlorphyll content in crop plants

2. Mepiquat chloride/Pix/PPC

Anti-gibberllin 1,1, dimethyl Piperidinium chloridae

Growth retardant, controls vegetative growth, increases chlorphyll synthesis and cause uniform maturity

3. Gibberlic acid Gibberellins Gibberellic acid Growth promoter, stimulates cell division, cell elongation, auxin metabolism, cell wall plasticity and permeability of cell membranes, RNA synthesis, induction of hydrolytic enzymes and increases plant height, increased mobilization and translocation of reserve food material

4. Miraculan Biological product containing tria contanol

Tria contanol (TRIA) the straight chain alcohol

Growth promoter has been found in increase the crop yield in enhancement in mobilization of photosynthetic activity and rapid increase in reducing sugars, soluble protein, succinate and changes in permeability of membrane

5. Cow urine consists of urea (10 to 30 mg/dL), uric acid, creatine amino acid and ammonia

Non-protein nitrogenous substance

Spraying the crop with cow and urine increase yield of aromatic grasses (Farmer’s note book, THE HINDU)

LEGEND

T1: NAA @ 20 ppm at 60 and 75 Days After Planting (DAP)

T2: NAA @ 40 ppm at 60 and 75 DAP

T3: Mepiquat chloride @ 500 ppm at 60 and 75 DAP

T4: Mepiquat chloride @ 1000 ppm at 60 and 75 DAP

T5: GA @ 20 ppm at 60 and 75 DAP

T6: GA @ 40 ppm at 60 and 75 DAP

T7: Miraculan @ 1000 ppm at 60 and 75 DAP

T8: Miraculan @ 2000 ppm at 60 and 75 DAP

T9: Cow urine (20% v/v) at 60 and 75 DAP

T10: Control

Fig.1: Plan of layout of experimental site

Plate1: General view of the patchouli experimental plot

3.6 Cultural operations 3.6.1 Land preparation

The land was prepared by deep ploughing and harrowing and the soil was brought to a fine tilth.

3.6.2 Planting

The 60 day old saplings raised from shoot cuttings were planted as per the recommended spacing 60 x 60 cm.

3.6.3 Fertilizer application

Recommended dose of fertilizer was 150 kg N, 50 kg P and 50 kg K. 50 kg N, 50 kg P and 50 kg K was applied at time of planting, remaining 100 kg N was applied in two split doses at 30 DAP and 60 DAP respectively.

3.6.4 After care

Irrigation was given at regular intervals. Endosulphan @ 1.5 ml l-1

at 30 DAS was sprayed to control leaf eating caterpillar.

3.6.5 Harvesting

The crop was harvested after 5 months of DAP, three plants from each plot were uprooted for yield parameters and herbage yield was recorded per plot and calculated on hectare basis.

Plate 2: Effect of plant regulators on morpho- physiological characters of patchouli

Plate2: Effect of plant regulators on morpho-physiological characters of patchouli

3.7 Collection of experimental data

Three plants were randomly selected from each plot and were tagged for recording various morphological observations at different stages.

3.7.1 Morphological parameters 3.7.1.1 Plant height

Plant height was recorded from base of the plant to the tip of main shoot of plant at 60, 90, 120 days after planting and at harvest. In each plot, three plants were selected and mean height was calculated and expressed in cm.

3.7.1.2 Number of leaves per plant

Total number of leaves was estimated by counting the leaf from tip to bottom of plant and is expressed as number per plant.

3.7.1.3 Number of branches/plant

The number of branches per plant was counted from three plants at 60, 90, 120 DAP and at harvest and mean was expressed as number of branches per plant.

3.7.2 Physiological and biochemical parameters 3.7.2.1 Chlorophyll content estimation

The chlorophyll content was estimated at 60, 90 and 120 DAP. Total chlorophyll, chlorophyll a and chlorophyll b contents were determined by following the method of Shoaf and Lium (1976). 100 mg of fresh leaf tissues were cut into small pieces and incubated in 7.0 ml of DMSO (dimethyl sulfoxide) at 65°C for 30 minutes. At the end of incubation period the supernatant was decanted and leaf tissues discarded. The volume was made up to 10 ml with DMSO. The absorbance was read at 645, 652 and 663 nm in UV-vis spectrophotometer (ELICO, 159). Total chlorophyll, chlorophyll a and chlorophyll b content were calculated using the formula given by Arnon (1949) and expressed in mg per gram fresh weight.

V

Chlorophyll 'a' = 12.7 (A663) - 2.69 (A645) x 1000 x W x a

V Chlorophyll 'b' = 22.9 (A645) - 4.68 (A663)x

1000 x W x a V Total chlorophyll = chlorophyll 'a' + chlorophyll 'b' = 27.8 (A652) x 1000 x W x a

Where,

A = Absorbance at specific wave length (645, 652, 663 nm) V = Final volume of the chlorophyll extract (ml) W = Fresh weight of the sample (g) a = Path length of light (1 cm)

3.7.2.2 Carotenoid estimation

The carotenoid content was estimated at 60, 90 and 120 days after planting following method of Goodwin and Briton (1988).

Carotene content was estimated in each treatment in all the three replications by

using acetone hexane extraction method. About 5 g of fresh leaf sample was taken from five leaves which was macerated by using 85 per cent acetone, till all the carotenoids are dissolved. The extract was filtered through muslin cloth and then transferred to 250 ml separating funnel. Then 100 ml distilled water and 100 ml hexane were added to the extract in the separating funnel. The solution was shaken properly and then allowed to settle. The lower layer of water was discarded and upper hexane layer was retained. The hexane fraction containing carotene was washed with distilled water for about three or four times with the help of separating funnel. The hexane fraction was then transferred to conical flask and to this smaller quantity of anhydrous sodium sulphate was added to remove the water completely in the final solution. Reading was taken at 448 nm wave length in the spectrophotometer.

3.7.2.3 Nitrate reductase activity

The nitrage reductase activity (NRA) in vivo was assayed at 60, 90 and 120 DAS following by the method of Sardhambal et al. (1978). Leaves were cut into small round disc, weighed and suspended in 0.1 M kNo3 under bright light for 1 hour for complete stomatal opening. Then discs were transferred to 25 ml volumetric flasks containing 5 ml of stock solution containing 0.1 M phosphate buffer (pH 7.5). 0.02 M kNo3, propanol (5%) and 2 drops of chloromphenicol (0.5 mg/ml). The flasks were incubated at 30°C for 30 minutes in dark and reaction was stopped by adding 0.1 ml of zinc acetate (0.1 M) and 1.9 ml of ethanol (70%). The contents were centrifuged at 3000 rpm for 10 minutes and supernatnat was collected. To the supernatent, 1.0 ml of sulphanilamide (1%) and 1 ml of NNEDA (N-Nipthyl ethylene diamine dihydrochloride 0.02%) were added and incubated at room temperature for 20 minutes. The activity of nitrate reductase was determined from a standard curve of kNo2 and

expressed as µmoles No2 formed per gram fresh weight per hour.

3.7.2.4 Total phenols

The phenol content was estimated at 60, 90 and 120 DAP by following Folin ciocatteau reagent form the alcohol filtrate (Sadasivam and Manikam, 1992) in oven dried samples.

Extraction

One gram of leaf tissue was weighed and made into small pieces and was boiled in 10 ml of 70 per cent alcohol for 5-10 minutes. The tissue was crushed with pestle and mortar and then filtered. The extracts were pooled and alcohol was evaporated on hot water bath and made up the volume to 10 ml with distilled water and this extract was stored in refrigerator at 4°C. Procedure

One ml alcohol extract was taken in a test tube to which one ml of FCR reagent followed by 2 ml of 2 per cent sodium carbonate was added. The tubes were shaken well and heated in a hot water bath for exactly one minute and then cooled under running tap water and blue colour developed was diluted with 25 ml distilled water and its absorbance was read at 650 nm in UV-vis spectrophotometer (ELICO, 159). The amount of phenols present in sample was calculated from a standard curve prepared from catechol and the content was expressed as mg per gram dry weight.

3.7.3 Anatomical characters

Number of oil glands was calculated by the replica method developed by Wolf et al. (1979) at 60, 90 and 120 DAP. Leaf samples were collected from three different plants in each treatment. Using applicator brush attached to correction fluid was dippled into thermocol, xylene flurol and then brush was stroked across the middle of the leaves, after a few seconds when the fluid became dry and firm replica was ready to be removed, a small piece of double adhesive celluloid tape was be recurred to a glass slide and the sticky surface was then placed over replica. The replica was mounted for viewing inprint of a distinct oil gland surrounded by large number of trichomes of dark structure. The member of oil glands were counted for three microscopic field and the mean of which was taken for further calculation. Then using occular and stage micrometer the area of microscopic field was calculated and number of oil glads per mm² was thus estimated.

3.7.4 Growth parameters 3.7.4.1 Total dry matter

The selected plant was uprooted from soil and washed to remove soil from the roots. The whole plant was divided into leaves, stem and roots and dried in oven for 15 hrs at 60°C and total weight of leaves and stem was weighed on balance in grams.

3.7.4.2 Leaf area

Leaf area was measured by leaf disc method as suggested by Vivekanandan et al. (1972). Twenty leaf discs of known size were taken using the cork borer from two plants. Both discs and leaves were oven dried at 75-80°C and leaf area was calculated by using the formula at different states of crop growth.

Wa x A LA = Wd

Where, LA = leaf area (dm²/plant) Wa = Weight of all leaves (inclusive of 20 discs weight) in grams Wd = Weight of discs (g) A = Area of disc

3.7.4.3 Leaf area index (LAI)

LAI was calculated by dividing the leaf area per plant by the land area occupied by the plant (Sestak et al., 1972).

Leaf area

LAI = Land area

3.7.4.4 Leaf area ratio (LAR)

The leaf area ratio was worked out by the formula of Radford (1967) and expressed as cm²/g.

Leaf area (cm2/plant)

LAR =

Total dry matter (g/plant)

3.7.4.5 Leaf area duration (LAD)

Leaf area duration is the integral of leaf area index over a growth period (Watson, 1952). LAD for various growth periods was worked out as per the formula of Power et al. (1967) and expressed in days.

L1 – L (i+1)

LAD = x (t2 - t1)

2

Where,

L1 = LAI at ith stage

L (i+1)= LAI at (i+1)th stage

t2-t1 = Time interval in days

3.7.4.6 Specific leaf area (SLA)

The inverse of specific leaf weight is the specific leaf area and was calculated as follows.

Leaf area (cm²)

SLA =

Leaf dry weight (g)

3.7.4.7 Specific leaf weight (SLW)

The specific leaf weight (g cm-2

) indicates the leaf thickness and was determined by the following formula.

Leaf dry weight (g) SLW = Leaf area (cm

2)

3.7.4.8 Absolute growth rate (AGR) (g plant

-1 day

-1)

It expresses the increasing dry weight per plant in unit time and was calculated by

using the following formula (Radford, 1967) and expressed as g plant-1

day-1

.

W2 - W1

AGR = t2 - t1

Where,

W1 = Total dry weight of the plant (g) at time t1

W2 = Total dry weight of the plant (g) at time t2 t2-t1 = Time interval in days

3.7.4.9 Relative growth rate

It is the rate of increase in dry weight (mg/g/day) already present and was calculated by using the formula of Blackman (1919).

loge W2 – loge W1

RGR =

t2-t1

3.7.4.10 Net assimilation rate (NAR)

Net assimilation rate is the rate of dry weight increased per unit leaf area per unit time. It was calculated by following the formula of Radford (1967) and expressed as mg cm

-2

day-1

W2 - W1 loge A2 – loge A1

NAR = x

t2 - t1 (A2-A1)

Where,

A1 and W1 = Leaf area (cm²) and total dry weight of plant (g) respective at time t1. A2 and W2 = Leaf area (cm²) and total dry weight of plant (g) respective at time t2. t2-t1 = Time interval in days

3.7.4.11 Crop growth rate (CGR)

Crop growth rate is the rate of dry matter production per unit ground area per unit time (Watson, 1952). It was calculated by using the following formula and expressed as g dm

-

2 day

-1.

W2 - W1 I

CGR = X t2 - t1 P

Where,

W1 = Dry weight of the plant (g) at time t1

W2 = Dry weight of the plant (g) at time t2 t2-t1 = Time interval in days P = Unit land area

3.7.5 Yield and yield components 3.7.5.1 Total herbage

The randomly selected three plants were uprooted in cool hours of morning from soil and the plants were washed to remove soil and leaves were separated and weighed and noted as fresh weight and leaves were shade dried by spreading in thin layer on hard dry surface in shed and after 60 days shade dry weight was calculated.

3.7.5.2 Percentage of essential oil content

The leaves were shade dried till they attained a constant shade dried weight and then the estimation of essential oil was patchouli is carried out using clevenger’s apparatus described by Guenther (1952) for 2-3 hours. Shade dried leaves were distilled for 6-12 hours, quantity of material used was 100 g in a 2000 ml round bottom flask filled ¾ with water. Essential oil got distilled along with steam vapours, on cooling in condenser, mixture of water and essential oil got collected in a graduated arm of clevenger’s apparatus. After formation of

separation of layer essential oil was measured and its percentage was calculated as v/w on on dry weight basis (DWB).

3.7.5.3 Essential oil yield/ha

The amount of oil obtained in mili litres was converted to kilograms by multiplying with specific gravity (0.95) (Shankaranarayan, 2002) of patchouli oil and expressed as kg/ha.

IV. EXPERIMENTAL RESULTS

An experiment was conducted at Main Agricultural Research Station, Dharwad to study the effect of plant regulators on growth and yield of patchouli during kharif season of the year 2004. The results of the experiment are presented in this chapter.

4.1 Morphological parameters 4.1.1 Plant height (cm) The plant height increased continuously from 60 days of transplanting (DAP) to harvest in the all the treatments. The experimental results on plant height revealed that treatments differed significantly at all the stages except at 60 DAP and it increased continuously from 60 DAP to harvest (Table 4). A maximum plant height (80.33) was obtained at 90 DAP in NAA @ 20 ppm. Among treatments GA3 @ 20 ppm (73.50), NAA @ 40 ppm (72.90) did not differ significantly with each other at 90 DAP and had higher plant height. Similarly, no significant differences were observed between different concentrations of miraculan, cow urine (20% v/v) and control. However, mepiquat chloride @ 500 ppm and @ 1000 ppm significantly recorded lower plant height as compared to control.

At 120 DAP, NAA @ 20 ppm continued to maintain higher plant height (84.31) which was almost on par with NAA @ 40 ppm (83.31) followed by GA3 @ 20 ppm which recorded plant height of 76.81 cm. However, GA3 @ 20 ppm was found to be on par with all the other treatments except mepiquat chloride @ 500 ppm and mepiquat chloride @ 1000 ppm which significantly recorded lower plant height compared to control. Similar trend was also noticed at harvest.

4.1.2 Number of branches The data on number of branches per plant as presented in table 5 indicated significant difference between the treatments except at 60 DAP. The number of branches significantly increased due to growth regulators at all the stages.

At 90 DAP, the treatment GA3 @ 20 ppm showed highest number of branches/plant (22.43) which is found to be on par with treatment GA3 @ 40 ppm (21.03 branches/plant) and also on par with NAA @ 40 ppm (20.03). No significant differences were obtained between different concentrations of miraculan, mepiquat chloride, cow urine (20% v/v) and NAA @ 20 ppm as compared to control.

At 120 DAP, the treatments GA3 @ 20 ppm and @ GA3 40 ppm recorded highest

number of branches/plant (32.03 and 29.90, respectively) followed by NAA @ 20 ppm which is on par with NAA @ 40 ppm. However, it is observed that miraculan @ 2000 ppm, cow urine (20% v/v) and control exhibited lower number of branches. Further, the lower number of branches were noticed in mepiquat chloride @ 1000 ppm which is on par with miraculan @ 1000 ppm.

The same trend was also noticed at harvest.

4.1.3 Number of leaves

It is evident from table 6 that number of leaves increased from 60 DAP to harvest. Significant differences in number of leaves per plant produced due to various treatments were observed at all the growth stages except at 60 DAP. At 90 DAP highest number of leaves per plant (528.0) was noticed in the treatment GA3 @ 20 ppm which is on par with NAA @ 20 ppm, GA3 @ 40 ppm and NAA @ 40 ppm and mepiquat chloride @ 500 ppm. No significant differences were observed between the treatments mepiquat chloride @ 1000 ppm, miraculan @ 2000 ppm and cow urine (20% v/v) as compared to control. Similar trend was noticed at

Table 4. Influence of plant growth regulators on plant height (cm) in patchouli

Days after planting Treatments

60 90 120 At harvest

T1 - NAA @ 20 ppm 27.26 80.33 84.31 88.86

T2 - NAA @ 40 ppm 28.00 72.90 83.31 88.10

T3 - Mepiquat chloride @ 500 ppm 27.96 57.43 60.19 63.20


T5 - GA @ 20 ppm 26.20 73.50 76.81 81.16

T6 - GA @ 40 ppm 26.16 69.00 75.81 81.13

T7 – Miraculan @ 1000 ppm 27.20 66.73 70.82 76.83

T8 – Miraculan @ 2000 ppm 28.26 62.90 72.90 75.26

T9 – Cow urine (20% v/v) 28.00 66.90 70.90 74.50

T10 – Control 27.50 64.56 70.21 75.36

Mean 27.31 67.06 72.31 76.22

S.Em± 0.670 2.455 2.399 2.57

CD at 5% NS 7.290 7.621 7.65

Table 5. Influence of plant growth regulators on number of branches in patchouli



T1 - NAA @ 20 ppm 3.2 18.70 26.10 30.13

T2 - NAA @ 40 ppm 3.9 20.03 25.10 29.16



T5 - GA @ 20 ppm 3.7 22.43 30.10 32.03

T6 - GA @ 40 ppm 3.4 21.03 29.90 31.73

T7 – Miraculan @ 1000 ppm 3.3 18.60 22.40 25.43

T8 – Miraculan @ 2000 ppm 3.2 18.60 23.20 25.2

T9 – Cow urine (20% v/v) 3.1 18.36 23.7 24.3

T10 – Control 3.2 18.30 23.3 24.6

Mean 3.46 19.34 25.16 28.05

S.Em± 0.38 0.670 1.22 1.26

CD at 5% NS 1.990 3.62 3.74

Table 6. Influence of plant growth regulators on number of leaves in patchouli



T1 - NAA @ 20 ppm 108.73 519.6 1160.60 1581.30

T2 - NAA @ 40 ppm 109.73 505.0 1091.60 1434.60



T5 - GA @ 20 ppm 112.36 528.0 1250.0 1761.60

T6 - GA @ 40 ppm 110.70 516.6 1153.30 1503.33

T7 – Miraculan @ 1000 ppm 110.36 414.0 1160.0 1164.00

T8 – Miraculan @ 2000 ppm 108.7 449.0 1035.0 1511.30

T9 – Cow urine (20% v/v) 108.7 456.6 1165.0 1471.60

T10 – Control 109.7 417.3 1034.0 1024.33

Mean 109.7 474.23 1104.0 1468.83

S.Em± 1.89 20.32 65.19 122.06

CD at 5% NS 60.36 193.61 362.51

Table 7. Influence of plant growth regulators on leaf dry weight (g/plant) in patchouli



T1 – NAA @ 20 ppm 4.93 69.92 108.16 131.50

T2 – NAA @ 40 ppm 4.33 59.50 91.40 105.20



T5 - GA @ 20 ppm 4.32 68.60 118.83 138.83

T6 - GA @ 40 ppm 4.30 59.50 101.90 125.20

T7 – Miraculan @ 1000 ppm 4.56 57.00 93.63 100.30

T8 – Miraculan @ 2000 ppm 4.43 56.21 73.56 104.90

T9 – Cow urine (20% v/v) 4.40 60.66 85.53 105.50

T10 – Control 4.76 43.60 66.56 89.90

Mean 4.70 59.28 59.98 109.68

S.Em± 0.495 4.54 4.864 4.509

CD at 5% NS 13.49 14.846 13.390

120 days after planting. At harvest the treatment GA3 @ 20 ppm recorded maximum number of leaves per plant (1761.6) which is on par with NAA @ 20 ppm, NAA @ 40 ppm and GA3 @ 40 ppm, miraculan @ 2000 ppm and cow urine. But, no significant differences were observed between treatments of different concentrations of mepiquat chloride miraculan @ 1000 ppm and cow urine (20% v/v). Control exhibited significantly lower number of leaves over all other treatments.

4.2 Growth parameters 4.2.1 Leaf dry weight (g/plant)

The leaf dry weight increased from 60 DAP to harvest in all the treatments and differed significantly in all the stages except at 60 DAP (Table 7). Among treatments, GA3 @ 20 ppm recoded maximum leaf dry weight (68.60 g) which is on par with all other treatments except treatments mepiquat chloride @ 1000 ppm and control which recorded lowest leaf dry weight (43.6 g) at 90 DAP. At 120 DAP GA3 @ 20 ppm showed highest leaf dry weight (118.83 g) which is on par with NAA @ 20 ppm. But, GA3 @ 40 ppm and NAA @ 20 ppm are on par with each other and no significant differences were observed among treatments NAA @ 40 ppm, mepiquat chloride @ 500 ppm, miraculan @ 1000 ppm and cow urine (20% v/v). Control recorded lowest leaf dry matter compared to all other treatments. At harvest GA3 @ 20 ppm recorded maximum leaf dry matter (138.83 g) which is on par with NAA @ 20 ppm and GA3 @ 40 ppm. No significant differences were observed in all other treatments. Control however recorded least leaf dry weight.

4.2.2 Stem dry weight (g/plant)

There was increase in the stem dry weight from 60 DAP to harvest (Table 8). Except 60 DAP the treatments showed significant differences at 90, 120 DAP and harvest. Among the treatments GA3 @ 20 ppm recorded significantly higher stem dry weight (54.86 g) followed by treatments NAA @ 20 ppm, GA3 @ 40 ppm, NAA @ 40 ppm. Cowurine (20% v/v) and which was on par with miraculan @ 2000 ppm and control @ 500 and 1000 ppm and miraculan 1000 ppm exhibited almost similar trend in the stem dry weight. At 120 DAP highest stem dry weight (117.06 g) was recorded by GA3 @ 20 ppm which was on par with treatments NAA @ 20 ppm and GA3 @ 40 ppm, where as all other remaining treatment failed to show significant difference in stem dry weight with control. At harvest, GA3 @ 20 ppm recorded highest stem dry weight (130.10 g) which is on par with treatments NAA @ 20 ppm and GA3 @ 40 ppm, whereas all other remaining treatments were on par with each other and control except NAA @ 40 ppm.

4.2.3 Total dry weight Total dry weight increased continuously from 60 DAP to harvest and treatments differed significantly at all the stages except at 60 DAP (Table 9). Among treatments, GA3 @ 20 ppm increased total dry weight (132.85 g) significantly followed by NAA @ 20 ppm, GA3 @ 20 ppm, NAA @ 40 ppm mepiquat chloride @ 500 ppm and cow urine (20% v/v) miraculan @ 1000 ppm and 2000 ppm exhibited higher stem dry weight than the control and mepiquat chloride @ 1000 ppm. Both the concentrations of miraculan, however were found to be on par with control. At 120 DAP significantly higher total dry matter (236.65 g) was recoded by GA3 @ 20 ppm which was found to be on par with NAA @ 40 ppm followed by all other remaining treatments which significantly increased total dry matter over the control except mepiquat chloride @ 1000 ppm which was on par with control. Similar trend was continued between the treatments at harvest.

Table 8. Influence of plant growth regulators on stem dry weight (g/plant) in patchouli



T1 - NAA @ 20 ppm 5.93 45.86 107.36 120.36

T2 - NAA @ 40 ppm 5.86 42.00 86.94 99.94



T5 - GA @ 20 ppm 6.06 54.86 117.06 130.10

T6 - GA @ 40 ppm 5.80 42.56 105.86 118.86

T7 – Miraculan @ 1000 ppm 5.80 37.93 74.46 88.86

T8 – Miraculan @ 2000 ppm 5.86 38.86 75.86 87.46

T9 – Cow urine (20% v/v) 5.90 40.86 73.30 85.23

T10 – Control 5.46 39.83 68.64 81.64

Mean 5.89 41.69 85.58 98.59

S.Em± 0.317 2.52 5.36 5.36

CD at 5% NS 7.494 15.94 15.93

Table 9. Influence of plant growth regulators on total dry matter (g/plant) in patchouli



T1 - NAA @ 20 ppm 10.37 115.20 216.23 252.61

T2 - NAA @ 40 ppm 10.40 100.34 179.09 189.92



T5 - GA @ 20 ppm 10.63 132.85 236.65 269.70

T6 - GA @ 40 ppm 10.31 102.27 208.45 205.78

T7 – Miraculan @ 1000 ppm 10.59 98.92 168.84 193.17

T8 – Miraculan @ 2000 ppm 10.49 95.85 150.22 191.17

T9 – Cow urine (20% v/v) 10.50 102.27 159.61 191.52

T10 – Control 10.45 84.20 135.94 172.29

Mean 10.73 101.22 176.30 209.02

S.Em± 0.59 5.711 7.01 7.34

CD at 5% NS 16.961 20.84 21.81

Table 10. Influence of plant growth regulators on leaf area (dm²/plant) in patchouli



T1 - NAA @ 20 ppm 8.77 32.89 52.18 63.75

T2 – NAA @ 40 ppm 7.73 28.09 42.80 48.46



T5 - GA @ 20 ppm 8.74 36.44 56.53 64.02

T6 - GA @ 40 ppm 7.29 30.62 56.07 62.44

T7 – Miraculan @ 1000 ppm 7.56 29.47 44.44 47.08

T8 – Miraculan @ 2000 ppm 8.34 29.89 32.67 46.19

T9 – Cow urine (20% v/v) 7.77 29.00 38.90 47.39

T10 – Control 7.64 19.97 31.48 41.95

Mean 8.36 28.93 43.25 51.69

S.Em± 1.087 1.968 2.742 2.667

CD at 5% NS 5.844 8.142 7.920

Table 11. Influence of plant growth regulators on leaf area index (LAI) in patchouli



T1 - NAA @ 20 ppm 0.24 0.91 1.46 1.73

T2 - NAA @ 40 ppm 0.21 0.75 1.18 1.34



T5 - GA @ 20 ppm 0.21 1.01 1.54 1.77

T6 - GA @ 40 ppm 0.20 0.81 1.44 1.77

T7 – Miraculan @ 1000 ppm 0.21 0.72 1.23 1.30

T8 – Miraculan @ 2000 ppm 0.23 0.73 1.39 1.28

T9 – Cow urine (20% v/v) 0.21 0.80 0.99 1.31

T10 – Control 0.21 0.55 0.88 1.16

Mean 0.22 0.77 1.22 1.43

S.Em± 0.03 0.060 0.07 0.074

CD at 5% NS 0.179 0.233 0.220

Table 12. Influence of plant growth regulators on leaf area duration (days) in patchouli

Treatments 60-90 90-120 120-150

T1 - NAA @ 20 ppm 17.36 35.44 48.54

T2 - NAA @ 40 ppm 15.09 29.44 46.29

T3 - Mepiquat chloride @ 500 ppm 14.59 28.95 36.26


T5 - GA @ 20 ppm 18.41 38.32 49.81

T6 - GA @ 40 ppm 15.43 32.15 47.54

T7 – Miraculan @ 1000 ppm 13.88 30.80 38.13

T8 – Miraculan @ 2000 ppm 14.54 24.68 32.86

T9 – Cow urine (20% v/v) 15.32 28.29 35.95

T10 – Control 11.51 24.77 30.59

Mean 5.08 29.79 40.43

S.Em± 1.098 1.917 3.846

CD at 5% 3.244 5.692 11.422

4.2.4 Leaf area (dm²) The data on leaf area presented in Table 10 indicated significant differences between the treatments at all the growth stages except at 60 DAP. At 90 DAP highest leaf area (36.44 dm²) was obtained in the treatment GA3 @ 20 ppm which was on par with NAA @ 20 ppm and GA3 @ 40 ppm followed by all remaining treatments which were on par with each other and recorded higher leaf area compared to control.

At 120 DAP GA3 @ 20 ppm continued to record significantly higher leaf area (56.53 dm²) which was on par with NAA @ 20 ppm and GA3 @ 40 ppm followed by NAA @ 40 ppm, miraculan @ 1000 ppm, mepiquat chloride @ 500 ppm which were on par with each other. But mepiquat chloride @ 1000 ppm, miraculan @ 2000 ppm and cow urine (20% v/v) did not show significant differences compared to control. A similar trend was continued between the treatments at harvest.

4.2.5 Leaf area index

It is evident from table 11 that the leaf area index was increasing from 60 DAP to harvest. The leaf area index recorded at 60 DAP showed non-significant results in all the treatments. At 90 DAP GA3 @ 20 ppm recorded significantly higher leaf area index (1.01) which was on par with NAA @ 20 ppm which were inturn on par with GA3 @ 40 ppm. And all other treatments significantly differed with each other except miraculan @ 1000 ppm which was on par with the control. A similar trend was continued between the treatments at 120 DAP.

At harvest GA3 @ 20 ppm recorded significantly higher LAI (1.77) which was on par with GA3 @ 40 ppm and NAA @ 20 ppm followed by all other treatments which failed to show significant differences with control.

4.2.6 Leaf area duration (days)

There was increase in leaf area duration form 60 DAP to harvest (Table 12). Among all treatments, significantly higher leaf area duration (18.41) was noticed in GA3 @ 20 ppm which was on par with NAA @ 20 ppm and GA3 @ 40 ppm followed by NAA @ 40 ppm and cow urine (20% v/v) and remaining treatments failed to yield significant difference among themselves and with control. During 90 to 120 DAP GA3 @ 20 ppm resulted significantly higher LAD (38.32) which was on par with NAA @ 20 ppm followed by NAA @ 40 ppm and GA3 @ 40 ppm which were on par with each other and both the concentrations of mepiquet chloride and miraculan used. Cow urine did not yield any significant difference over control.

At harvest significantly higher LAD was obtained (49.81) in GA3 @ 20 ppm which was

on par with GA3 @ 40 ppm and NAA @ 20 ppm and NAA @ 40 ppm followed by all other treatments having no significant difference among each other and were on par with control.

4.2.7 Leaf area ratio (cm²/gm)

It is evident form Table 13 that leaf area ratio was highest at 60 DAP, it declined considerably at 90 and 120 DAP, there after remained constant at harvest. No significant difference was observed among the treatments with respect to leaf area ratio at all stages of crop.

Table 13. Influence of plant growth regulators on leaf area ratio (cm²/g/plant) in patchouli



T1 - NAA @ 20 ppm 79.16 28.59 24.28 24.01

T2 - NAA @ 40 ppm 74.87 26.72 23.91 23.56



T5 - GA @ 20 ppm 72.92 27.43 23.47 22.42

T6 - GA @ 40 ppm 70.50 25.89 25.41 25.15

T7 – Miraculan @ 1000 ppm 71.51 29.67 26.23 24.71

T8 – Miraculan @ 2000 ppm 79.70 27.65 24.93 23.93

T9 – Cow urine (20% v/v) 74.21 28.32 24.36 23.93

T10 – Control 73.80 27.08 23.19 22.02

Mean 75.97 27.78 24.67 25.07

S.Em± 6.96 0.74 1.502 1.682

CD at 5% NS NS NS NS

Table 14. Influence of plant growth regulators on specific leaf area (cm²/g) in patchouli



T1 – NAA @ 20 ppm 48.17 48.83 48.17 47.51

T2 - NAA @ 40 ppm 6.78 47.15 46.78 45.53



T5 - GA @ 20 ppm 52.46 49.29 52.46 46.08

T6 - GA @ 40 ppm 51.77 47.94 51.77 51.01

T7 – Miraculan @ 1000 ppm 44.68 45.67 47.41 46.78

T8 – Miraculan @ 2000 ppm 47.41 45.94 44.68 44.12

T9 – Cow urine (20% v/v) 47.31 47.92 45.57 44.98

T10 – Control 45.57 45.88 47.31 46.68

Mean 47.73 47.32 47.73 47.03

S.Em± 1.697 0.698 1.679 1.624

CD at 5% NS 2.073 NS NS

Table 15. Influence of plant growth regulators on specific leaf weight (mg/cm²) in patchouli



T1 - NAA @ 20 ppm 5.56 21.79 20.75 21.04

T2 - NAA @ 40 ppm 5.65 20.87 21.42 21.70



T5 - GA @ 20 ppm 5.65 21.86 22.45 21.79

T6 - GA @ 40 ppm 5.94 21.23 21.13 21.70

T7 – Miraculan @ 1000 ppm 6.03 20.85 21.42 21.41

T8 – Miraculan @ 2000 ppm 5.37 20.47 21.98 22.74

T9 – Cow urine (20% v/v) 5.65 21.79 21.12 22.26

T10 – Control 6.22 20.26 21.13 21.42

Mean 5.75 21.13 21.04 21.32

S.Em± 0.42 0.31 0.69 0.690

CD at 5% NS 0.92 NS NS

4.2.8 Specific leaf area (cm²/gm)

The information on specific leaf area as influenced by various treatments during different growth periods are furnished in Table 14 which revealed that no significant difference was observed among the treatments with respect to specific leaf area in all the growth stages except 90 DAP where GA3 @ 20 ppm recorded significantly higher SLA (49.29) which was on par with NAA @ 40 ppm, cow urine (20% v/v) and GA3 @ 40 ppm followed by other treatments which yielded no significant difference over control.

4.2.9 Specific leaf weight (mg/cm²)

Specific leaf weight (SLW) as influenced by the application of different growth regulators is presented in Table 15. There was increase in SLW from 60 DAP to 90 DAP, thereafter it remained static till harvest in all the treatments. Treatments differed significantly only at 90 DAP and no significant difference was observed with treatments at 60 DAP, 120 DAP and at harvest. Among treatments GA3 @ 20 ppm recorded significantly higher SLW (21.86) over control which was on par with all treatments except both the concentrations of miraculan i.e. @ 1000 and 2000 ppm which showed no significant difference with control.

4.2.10 Absolute growth rate (g/plant/day)

Absolute growth rate decreased continuously from 60 DAP to harvest in all treatments and treatments differed significantly in all the stages (Table 16).

Maximum AGR was noticed in GA3 @ 20 ppm (4.070) followed by NAA @ 20 ppm,

NAA @ 40 ppm, GA3 @ 40 ppm and cow urine (20% v/v) which were on par with each other. No significant difference was obtained between 60 to 90 DAP, different concentrations of mepiquat chloride and miraculan used were on par with control between 60 to 90 DAP.

AGR between 90-120 DAP was significantly higher in GA3 @ 20 ppm which was on

par with NAA @ 20 ppm this was followed by NAA @ 40 ppm, GA3 @ 40 ppm, mepiquat chloride @ 500 ppm which were on par with each other. However, treatments mepiquat chloride @ 1000 ppm, miraculan @ 2000 ppm and cow urine (20% v/v) did not yield significant difference compared to control. The same trend was noticed between the treatment between 120 DAP and harvest.

4.2.11 Crop growth rate (g/dm²/day)

Information on crop growth rate as influenced by various treatments during different growth periods are furnished in Table 17. In general CGR recoded maximum at 60-90 DAP and thereafter it declined at 90-120 DAP and 120 DAP to harvest except GA3 treatment in which slight increase in the crop growth rate was observed at 90-120 DAP, but it declined thereafter. Maximum CGR (0.096) was observed in GA3 @ 20 ppm followed by NAA @ 20 ppm, GA3 @ 40 ppm, NAA @ 20 pm and cow urine (20% v/v) which were on par with each other and no significant difference was obtained between different concentration of mepiquat chloride and effluent concentrations of miraculan in comparison with control. Between 90-120 DAP the CGR was significantly higher in GA3 @ 20 ppm which was on par with NAA @ 20 ppm and GA3 @ 40 ppm followed by NAA @ 40 ppm, mepiquat chloride @ 500 ppm and miraculan @ 1000 ppm which were on par with each other. But, cow urine (20% v/v), miraculan @ 2000 ppm and mepiquat chloride @ 1000 ppm recorded no significant difference with control. This same trend was also noticed between treatments between 120 DAP and harvest.

4.2.12 Relative growth rate (mg/g/day)

The data on relative growth rate as influenced by the growth regulators is presented in Table 18 which indicated that RGR was higher at 60-90 DAP and then declined continuously at 90-120 DAP and 120 DAP to harvest in all the treatments.

Table 16. Influence of plant growth regulators on absolute growth rate (AGR, g/day) in patchouli

Treatments 60-90 90-120 120-150

T1 - NAA @ 20 ppm 3.467 3.363 1.428

T2 - NAA @ 40 ppm 2.997 2.223 0.887



T5 - GA @ 20 ppm 4.070 3.457 1.430

T6 - GA @ 40 ppm 3.057 2.597 1.207

T7 – Miraculan @ 1000 ppm 2.940 2.323 1.099

T8 – Miraculan @ 2000 ppm 2.840 1.810 1.428

T9 – Cow urine (20% v/v) 3.053 1.907 1.061

T10 – Control 2.453 1.720 1.210

Mean 3.008 2.465 1.088

S.Em± 0.184 0.150 0.057

CD at 5% 0.546 0.446 0.170

Table 17. Influence of plant growth regulators on crop growth rate (CGR, g/dm²/day) in patchouli

Treatments 60-90 90-120 120-150

T1 - NAA @ 20 ppm 0.085 0.096 0.033

T2 - NAA @ 40 ppm 0.082 0.071 0.030



T5 - GA @ 20 ppm 0.096 0.099 0.039

T6 - GA @ 40 ppm 0.083 0.093 0.033

T7 – Miraculan @ 1000 ppm 0.082 0.064 0.019

T8 – Miraculan @ 2000 ppm 0.079 0.050 0.039

T9 – Cow urine (20% v/v) 0.085 0.053 0.029

T10 – Control 0.068 0.048 0.035

Mean 0.080 0.069 0.030

S.Em± 0.003 0.003 0.001

CD at 5% 0.009 0.008 0.003

Table 18. Influence of plant growth regulators on relative growth rate (RGR, g/g/day) in patchouli

Treatments 60-90 90-120 120-150

T1 – NAA @ 20 ppm 33 09 03

T2 – NAA @ 40 ppm 32 08 02

T3 – Mepiquat chloride @ 500 ppm 30 07 02

T4 – Mepiquat chloride @ 1000 ppm 28 07 02

T5 - GA @ 20 ppm 36 10 03

T6 - GA @ 40 ppm 32 08 03

T7 – Miraculan @ 1000 ppm 32 07 02

T8 – Miraculan @ 2000 ppm 31 06 02

T9 – Cow urine (20% v/v) 32 06 02

T10 – Control 29 07 02

Mean 32 0.75 0.23

S.Em± 0.1 0.01 0.01

CD at 5% 0.3 0.1 NS

Table 19. Influence of plant growth regulators on net assimilation rate (NAR, mg/cm²/day) in patchouli

Treatments 60-90 90-120 120-150

T1 - NAA @ 20 ppm 0.888 0.350 0.144

T2 - NAA @ 40 ppm 0.827 0.325 0.081



T5 - GA @ 20 ppm 0.957 0.416 0.161

T6 - GA @ 40 ppm 0.857 0.344 0.141

T7 – Miraculan @ 1000 ppm 0.799 0.305 0.083

T8 – Miraculan @ 2000 ppm 0.786 0.278 0.081

T9 – Cow urine (20% v/v) 0.822 0.217 0.061

T10 – Control 0.845 0.263 0.067

Mean 0.822 0.310 0.089

S.Em± 0.021 0.011 0.005

CD at 5% 0.063 0.033 0.014

RGR showed no significant difference between 120 DAP to harvest. At 60-90 DAP the RGR recorded significantly higher value (0.36) in GA3 @ 20 ppm

which was on par with NAA @ 20 ppm followed by all other treatments except different concentration of mepiquat chloride which were on par with control.

The same trend was noticed among the treatments between 90-120 DAP.

4.2.13 Net assimilation rate (mg/cm²/day)

It is evident from Table 19 that the net assimilation rate was higher at 60-90 DAP and then declined at 90-120 DAP and 120 to harvest in all the treatments. Among the treatments between 60-90 DAP GA3 @ 20 ppm recorded maximum NAR (0.957) followed by NAA @ 20 ppm, GA3 @ 40 ppm and NAA @ 40 ppm and cow urine (20% v/v) which were found to be on par with each other and control. Between 90-120 DAP NAR was significantly high (0.410) at GA3 @ 20 ppm followed by NAA @ 20 ppm, GA3 @ 40 ppm and NAA @ 40 ppm. No significant difference was observed between remaining treatments which were on par with each other and control. The similar trend was also noticed between 120 DAP to harvest.

4.3 Anatomical parameter 4.3.1 Number of oil glands (per mm² leaf area) The data on number of oil glands as influenced by growth regulators is presented in Table 20 which revealed that on an average irrespective of the treatment, no significant difference was observed at 60 DAP. Whereas at 90 DAP and 120 DAP NAA @ 20 ppm recoded significantly higher number of oil glands (21.11 and 21.88 respectively) which was on par with GA3 @ 20 ppm, NAA @ 40 ppm and GA3 @ 40 ppm followed by both the concentrations of mepiquat chloride and miraculanused and cow urine (20% v/v) which were on par with each other and control.

4.4 Biochemical parameters 4.4.1 Chlorophyll ‘a’ and chlorophyll ‘b’ content (mg/g fresh weight)

It is evident from Table 21 that the chlorophyll ‘a’ content did not differ significantly between treatments at 60 DAP and 120 DAP. At 90 DAP significantly higher chlorophyll ‘a’ content (1.400) was obtained in GA3 @ 20 ppm which was on par with GA3 @ 40 ppm followed by all treatmetns which were on par with control except the two concentrations of mepiquat chloride i.e. @ 1500 and 1000 ppm respectively which recorded significantly lower chlorophyll content than control.

No significant differences were observed among the treatments with respect to chlorophyll ‘b’ content in leaves at any of the growth stages (Table 21).

4.4.2 Total chlorophyll (mg/g fresh weight) Influence of plant growth regulators indicated no significant differences on total chlorophyll at 60 DAP and 120 DAP (Table 22) where as at 90 DAP, significantly higher total chlorophyll content (2.116) was obtained with GA3 @ 20 ppm which was on par with GA3 @ 40 ppm followed by all treatments which were on par with each other and control except mepiquat chloride @ 1000 ppm.

Table 19. Influence of plant growth regulators on net assimilation rate (NAR, mg/cm²/day) in patchouli

Treatments 60-90 90-120 120-150

T1 - NAA @ 20 ppm 0.888 0.350 0.144

T2 - NAA @ 40 ppm 0.827 0.325 0.081



T5 - GA @ 20 ppm 0.957 0.416 0.161

T6 - GA @ 40 ppm 0.857 0.344 0.141

T7 – Miraculan @ 1000 ppm 0.799 0.305 0.083

T8 – Miraculan @ 2000 ppm 0.786 0.278 0.081

T9 – Cow urine (20% v/v) 0.822 0.217 0.061

T10 – Control 0.845 0.263 0.067

Mean 0.822 0.310 0.089

S.Em± 0.021 0.011 0.005

CD at 5% 0.063 0.033 0.014

Table 21. Influence of plant growth regulators on chlorophyll ‘a’ and ‘b’ content (mg/g fresh weight) in patchouli

Chlorophyll ‘a’ Chlorophyll ‘b’

Days after planting Days after planting Treatments

60 90 120 60 90 120

T1 - NAA @ 20 ppm 1.303 1.322 1.303 0.6993 0.7017 0.7007

T2 - NAA @ 40 ppm 1.300 1.249 1.300 0.6993 0.7013 0.6993

T3 - Mepiquat chloride @ 500 ppm 1.303 1.247 1.293 0.6997 0.6987 0.6980

T4 - Mepiquat chloride @ 1000 ppm 1.293 1.250 1.303 0.7010 0.6980 0.6987

T5 - GA @ 20 ppm 1.303 1.400 1.303 0.7007 0.7000 0.6993

T6 - GA @ 40 ppm 1.290 1.371 1.290 0.7007 0.6970 0.7007

T7 – Miraculan @ 1000 ppm 1.303 1.307 1.300 0.6983 0.6997 0.6973

T8 – Miraculan @ 2000 ppm 1.300 1.309 1.303 0.6973 0.7000 0.6983

T9 – Cow urine (20% v/v) 1.300 1.312 1.310 0.6973 0.696 0.6993

T10 – Control 1.310 1.308 1.300 0.6993 0.6963 0.6973

Mean 1.302 1.307 1.302 0.6993 0.6989 0.6993

S.Em± 0.347 0.0144 0.347 0.027 0.017 0.027

CD at 5% NS 0.0429 NS NS NS NS

Table 22. Influence of plant growth regulators on total chlorophyll (mg/g fresh weight) in patchouli


60 90 120

T1 – NAA @ 20 ppm 2.02 2.023 1.994

T2 - NAA @ 40 ppm 1.999 1.984 1.999



T5 - GA @ 20 ppm 1.994 2.116 2.002

T6 - GA @ 40 ppm 2.003 2.091 2.003

T7 – Miraculan @ 1000 ppm 1.988 2.007 2.003

T8 – Miraculan @ 2000 ppm 2.000 2.043 1.988

T9 – Cow urine (20% v/v) 1.999 2.008 2.073

T10 – Control 2.073 2.004 1.999

Mean 2.010 2.020 2.010

S.Em± 0.338 0.166 0.338

CD at 5% NS 0.493 NS

Table 23. Influence of plant growth regulators on carotenoid content (mg/100 g fresh weight in patchouli


60 90 120

T1 – NAA @ 20 ppm 10.37 13.33 13.55

T2 - NAA @ 40 ppm 10.40 13.55 13.55

T3 – Mepiquat chloride @ 500 ppm 11.67 13.55 13.61


T5 - GA @ 20 ppm 10.63 13.44 13.81

T6 - GA @ 40 ppm 10.31 13.77 13.78

T7 – Miraculan @ 1000 ppm 10.59 13.78 13.22

T8 – Miraculan @ 2000 ppm 10.49 13.22 14.11

T9 – Cow urine (20% v/v) 10.50 14.00 13.38

T10 – Control 10.45 13.88 13.62

Mean 10.73 13.62 13.21

S.Em± 0.59 0.78 0.78

CD at 5% NS NS NS

4.4.3 Carotenoid content (mg/100 g fresh weight) Observations on carotenoid content as influenced by various growth regulators are presented in Table 23. Carotenoid content was not affected by growth regulator treatments at any of the stages where in no significant difference was observed among treatments at any of growth stage.

4.4.4 Phenol content (mg/g dry wt.)

Observations on phenol content as influenced by various growth regulators are presented in Table 24. The phenol content increased continuously from 60 DAP to 120 DAP. No significant difference was observed among treatments at 60 DAP. At 90 DAP the treatment NAA @ 20 ppm recorded significantly higher phenol content (0.533) followed by GA3 @ 20 ppm, NAA @ 40 ppm and both the concentrations of mepiquat chloride which were on par with each other where as miraculan @ 1000 and 2000 pm and cow urine failed to show significant difference with control. The similar trend was noticed between treatments at 120 DAP.

4.4.5 Nitrate reductase activity (µmoles NO2 fixed/g fresh weight/ hour)

The data on nitrate reductase activity as influenced by growth regulators is presented in Table 25. The results revealed that NRA was highest during 60 DAP and declined thereafter at 90 and 120 DAP.

No significant difference was observed among the treatments at 60 DAP where as at

90 DAP GA3 @ 20 ppm recorded significantly higher NRA (48.70) which was on par with NAA @ 20 ppm. GA3 @ 40 PPM followed by remaining treatments which were on par with each other and control.

At 120 DAP NRA was significantly higher (29.23) in treatment GA3 @ 20 ppm which

was on par with NAA @ 20 ppm and GA3 @ 40 ppm this is followed by mepiquat chloride @ 500 ppm and NAA @ 40 ppm which were on par with each other. Mepiquat chloride @ 1000 ppm, miraculan @ 2000 ppm and cow urine (20% v/v) did not show significant different among each other and also over control.

4.5 Yield parameters 4.5.1 Fresh weight (q/ha) of leaves

Fresh weight of leaves showed significant differences among the treatments at harvest (Table 26). Significantly higher fresh weight (53.62) was obtained in treatment GA3 @ 20 ppm which was on par with NAA @ 20 ppm and GA3 @ 40 ppm followed by NAA @ 40 ppm and miraculan @ 1000 ppm. All other treatments failed to yield significant difference with control. In general, all the treatments had more fresh weight over control.

4.5.2 Shade dry weight (q/ha) of leaves

The data on shade dry weight of leaves presented in Table 26. Indicated a significant difference among treatments. At harvest treatment GA3 @ 20 ppm recorded significantly higher (13.07) shade dry weight which was on par with NAA @ 20 ppm GA3 @ 40 ppm followed by NAA @ 40 ppm and miraculan @ 1000 ppm. Remaining treatments did not have significant difference over control. All treatments in general had more shade dry weight over control.

Table 24. Influence of plant growth regulators on phenols content (mg per g dry weight) in patchouli


60 90 120

T1 – NAA @ 20 ppm 0.260 0.533 0.740

T2 - NAA @ 40 ppm 0.267 0.467 0.677



T5 - GA @ 20 ppm 0.250 0.423 0.640

T6 - GA @ 40 ppm 0.247 0.320 0.570

T7 – Miraculan @ 1000 ppm 0.253 0.290 0.513

T8 – Miraculan @ 2000 ppm 0.240 0.310 0.553

T9 – Cow urine (20% v/v) 0.247 0.293 0.487

T10 – Control 0.257 0.293 0.450

Mean 0.252 0.378 0.585

S.Em± 0.012 0.015 0.017

CD at 5% NS 0.044 0.052

Table 25. Influence of plant growth regulators on nitrate reductase activity NRA (µµµµ moles of NO2 fixed/g fresh weight) in patchouli


60 90 120

T1 - NAA @ 20 ppm 68.06 47.43 28.76

T2 - NAA @ 40 ppm 64.73 44.50 24.50


T4 - Mepiquat chloride @ 1000 ppm 64.36 4.2.43 22.26

T5 - GA @ 20 ppm 63.03 48.70 29.23

T6 - GA @ 40 ppm 64.3 45.83 27.53

T7 – Miraculan @ 1000 ppm 61.20 45.53 28.40

T8 – Miraculan @ 2000 ppm 66.23 44.03 23.93

T9 – Cow urine (20% v/v) 67.46 43.10 22.76

T10 – Control 66.03 42.10 21.43

Mean 64.86 44.60 24.83

S.Em± 1.551 1.220 0.801

CD at 5% NS 3.622 2.380

4.5.3 Oil percentage (%)

Results revealed that treatments differed significantly in oil per cent (Table 26). A maximum oil recovery per cent of 2.98 was recorded in NAA @ 20 ppm followed by GA3 @ 20 ppm and NAA @ 40 ppm, GA3 @ 40 ppm which were on par with each other. Significant differences with regard to oil percentage was not obtained between the treatments, cow urine (20% v/v) and control. However all the treatments in general increased oil percentage except cow urine.

4.5.4 Oil content (kg/ha)

It is evident from Table 26 that the oil content showed significant difference among treatments at harvest. Maximum oil content (37.00) was obtained in GA3 @ 20 ppm which was on par with NAA @ 20 ppm followed by GA3 @ 40 ppm and remaining all other treatments showed significant increase in oil content over control except cow urine (20% v/v) which was on par with control. All treatments in general increased the oil content except cow urine (20% v/v) over control.

Table 26. Influence of plant growth regulators on yield parameters of patchouli (FWB and DWB)

Herbage yield

Treatments Fresh weight basis (q/ha)

Shade dry weight basis (q/ha)

Oil (%) Oil content (kg/ha)

T1 - NAA @ 20 ppm 51.59 12.55 2.98 35.56

T2 - NAA @ 40 ppm 47.70 11.52 2.82 29.05



T5 - GA @ 20 ppm 53.62 13.07 2.84 37.00

T6 - GA @ 40 ppm 49.16 12.04 2.92 33.39

T7 – Miraculan @ 1000 ppm 47.85 11.63 2.53 27.74

T8 – Miraculan @ 2000 ppm 44.67 10.86 2.52 26.36

T9 – Cow urine (20% v/v) 42.81 10.58 2.47 24.85

T10 – Control 4226 10.29 2.49 24.36

Mean 47.09 11.52 2.67 29.36

S.Em± 1.609 0.506 0.001 0.808

CD at 5% 4.778 1.502 0.002 2.404

V. DISCUSSION

Plant growth and development is a complex process. There are two important factors which influence the reaction and metabolism of plants and thus regulate the developmental pattern of a plant, one of them is a system of endogenous chemical messengers, called hormones which exert important regulatory effects upon the development of individual organs of the plant as well as the plant as a whole. The second one comprises more or less interdependent set of external environmental factors, such as light, water temperature and gravity, which play indispensable role in the development as do hereditary factors which have been transmitted to it from its biological parents. Thus, the final pattern of development and behaviour of each individual plant is the remit of a complex interplay between genetic, hormonal and environmental factors.

The role of plant growth regulators in optimising the yield and quality of various

economical parts of crop plants is an established fact. PGRs are known to modify growth by increasing or decreasing the morphological traits such as plant height, number of branches etc. The plant growth regulators optimise the yield of the plant by bringing necessary physiological changes. Patchouli being an aromatic crop contains the essential oil in its leaves, stem and also flowers in some species. It is possible to increase the herbage and also the oil yield by exogenous application of PGRs.

The plant hormones are broadly been characterized into five distinct and diversified

groups. These include auxins and gibberllins, that stimulate predominantly cell elongation the cytokinins which are purine bases that stimulate cell division, ethylene, the olefinic gaseous molecule that regulates fruit ripening and abscisic acid (ABA), the sesquiterpene that regulates senescence and abscission of plant parts and helps in plant water relations. While these bioregulators are distinctive both in chemical characteristics and in exhibiting characteristic growth responses, each of the bio-regulators has potentiality to alter almost all the aspects of plant growth and development as both promoter as well as retardant when used in appropriate concentrations.

Although extensive studies on the influence of PGRs on various crop plants have

been made the studies on the effect of PGRs on aromatic and medicinal plants is very much limited. Therefore, the present study was undertaken to study the effect of growth regulators (promoters and retardants) on growth and yield of patchouli and results obtained are discussed in this chapter.

5.1 Morphological parameters

Growth regulators significantly influence the morphological characters such as plant height, number of branches and number of leaves per plant.

Basically plant height is a genetically controled character. But, several studies have

indicated that plant height can be either increased or decreased by application of synthetic plant growth regulators. However, in the present investigation a significant difference in plant height was noticed among the treatments by the application of different plant growth regulators used. It is interesting to note that there was an increase in plant height over control in all the treatments except the two concentrations of mepiquat chloride used in the study which decreased plant height.

The plant height was significantly higher with NAA @ 20 ppm followed by GA3 @ 20

ppm. This clearly indicated that the mode of action of these growth regulators vary from one type to another and one concentration to another concentration.

The increased plant height by NAA application might be due to primary role of auxins

viz., cell division, cell elongation there by high deposition of cell wall material through catalysing activities of carboxylase and peptidase and also by inhibition of antagonistic effect of inhibitors particularly abscession as reported by Pandey (1975) and reduction in plant

height due to mepiquat chloride appears to be because of reduction in cell division and cell elongation which caused inhibitory action in the biosynthetic pathway of GA3 (Moore, 1980).

Similar observations were also reported by Bhaskar et al. (1997) and Misra (1995) in

patchouli. Duhan et al. (1978) also reported same in Mentha citrata. The number of leaves and number of branches per plant are important morphological

characters which are directly related to herbage yield. The application of growth regulators increased the number of leaves and total number of branches per plant. The data on the number of leaves and total number of branches revealed that the growth regulators showed a significant increase in number of leaves and total number of branches in the present investigation at 90, 120 DAP and harvest. The treatment GA3 @ 20 ppm and NAA @ 20 ppm recorded significantly higher number of leaves and number of branches as compared to control. Similarly Krishnamoorthy and Madalageri (2000) and Mousa and Emary (1983) reported increased number of leaves and number of branches by application of growth regulators in ajowan and patchouli respectively. The increased number of branches per plants perhaps is due to increased polar transport of auxins towards basal axillary buds leading to increased number of branches, thereby increasing the number of leaves by growth regulators and this may be attributed to production of secondary and teritary branches (Krishnamoorthy and Madalageri, 2000).

5.2 Dry matter production

Poor translocation of photo assimilates to the growing parts is the major constraint in many of crops. This constraint can be overcome by applying synthetic plant growth regulators which improve the canopy structure and increase the productivity through the manipulation of source sink relationship.

The dry matter accumulation in the leaf increased up to harvest in all the growth

regulator treatments. The leaf dry weight was significantly higher with the application of GA3 @ 20 ppm followed by NAA @ 20 ppm which were found on par with each other.

The higher leaf dry weight may be due to higher number of leaves per plant. Similar

results were also by Krishnamoorthy and madalageri (2000) in ajowan and Bhattacharya et al. (1995) in geranium.

Similarly stem dry weight also increased significantly due to the application of growth

regulators GA3 @ 20 ppm, NAA @ 20 ppm and GA3 @ 40 ppm at 90, 120 DAP and harvest as compared to control.

The amount of total dry matter produced is an indication of the overall efficiency of

utilization of resources and better light interception. The data pertaining to total dry weight per plant indicated that, it increased continuously from 60 DAP to harvest. At later stage of crop growth, the dry matter accumulated at a decreasing rate.

Total dry weight of plant at harvest was significantly influenced by GA3 @ 20 ppm

followed by NAA @ 20 ppm compared to control. This increase in dry weight of the plant could be attributed to increase in plant height, number of branches and number of leaves per plant. In addition it was also observed that there was significant increase in LAI, LAD, LAR, SLA, SLW and biochemical parameters such as chlorophyll, NRA activity together might have influenced for enhanced total dry matter. These results are in confirmation with the findings of Umesha et al. (1991) who reported that maximum amount of dry matter was noticed in clocimum due to application of Gibberlic acid. Similar results were reported by Doijode (1975) in pea, and Deotole et al. (1998) in safflower.

5.3 Growth parameters

In the present study leaf area (LA), leaf area index (LAI) and leaf area duration increased up to harvest and in general application of growth regulators showed a profound effect over these parameters and significant differences were noticed among the growth regulator treatments at all stages except at 60 DAP where in GA3 @ 20 ppm and NAA @ 40 ppm recorded highly significant leaf area, leaf area index and leaf area duration as compared to control at 90, 120 DAP and harvest. However, leaf area ratio failed to show any significant differences among treatments at any of growth stages.

The increased leaf area and leaf area dependent components like LAI and LAD

because of GA3 application might be due to the fact that enhances cell division by promoting DNA synthesis. The application of gibberellin to most growing plant is the increase in length of stems, primary cause for which is increased length of the cells. But, I is also mentioned that in some plants increase is also due to cell division often accompanied by an increase in cell size and finally concluded that depending upon the plant increased size is either due to increase in cell division or cell elongation or both. Further, Bhattacharjee (1993) stated GA3 promotes uptake at Ca

++ ions into cytoplasm and increases cell wall extensively. Another two

parameters determined by leaf area is LAI and LAD. The LAI and LAD was significantly higher with growth promoters, GA3 @ 20 ppm and NAA @ 20 ppm. This could be attributed to retention of green leaves for longer duration and higher leaf area index (Bhattacharjee, 1993).

Growth promoting substances used in the present experiment namely GA3 and NAA

had a positive effect on cell division and cell elongation leading to enhanced leaf expansion causing maintenance of more green leaf area especially during later phases of growth and development.

This is in accordance with Singh (2003a) who reported that foliar application of GA3

(100 ppm) increased leaf area in french marigold. Umesha et al. (1991) also reported that with the foliar application of GA, there was increased leaf area in clocimum.

Specific leaf area and specific leaf weight were another two parameters influenced by

growth regulators treatments only at 90 DAP and at later stages treatments failed to impart significant differences in SLA and SLW. At 90 DAP GA3 @ 20 ppm recorded highest SLA and SLW. This indicates that GA3 application might have increased growth by altering dry matter distribution to increase total leaf area or specific leaf area by increasing photosynthetic rate per unit area of leaf which is also in agreement with Lester et al. (1972). Similar observations were also made by Bugbee and John (1984) who reported increased SLA in Tomato plants treated with GA3 and Gupta et al. (1995) reported increased SLW in Ocimum carnosum where growth regulators were applied.

The growth analysis parameter AGR indicated significant differences due to growth

regulator treatments at all the stages. In general AGR was highest at 60-90 DAP and decreased thereafter towards harvest. AGR was significantly higher in treatment GA3 @ 20 ppm at all the stages. The results are in confirmation with findings of Patil et al. (1985) who reported higher AGR in treatment GA3 125 ppm compared to rest of treatments in chilli. Kid et

al. (1993) also reported increased AGR due to application of TIBA in soybean. The crop growth rate (CGR) is also an important factor influenced by LAI,

photosynthetic rate and leaf angle and is an index of amount of radiation energy intercepted. CGR was highest between 60-90 DAP thereafter it declined towards harvest. The growth regulators significantly increased the CGR at 60-90, 90-120 DAP and at harvest. The present data also revealed that application of GA3 @ 20 ppm resulted in increased CGR especially at 90-120 DAP and it was due to increased dry matter partitioning in stem and leaves. Similarly, Gupta et al. (1995) observed increased CGR due to the application of growth regulator (Triacontanol) in Ocimum carnosum.

The relative growth rate (RGR) is also an important growth parameter which decreased from 60 DAP to harvest. The RGR showed significant differences due to growth regulator treatment at 60-90 DAP, 90-120 DAP but failed to impart significant difference between 120 DAP to harvest.

Shrivastava and Tiwari (1981) indicated that spraying of TIBA and planofix

significantly increased RGR in chickpea. Similarly, the present data also indicated that during early stage of crop growth, RGR was significantly more with use of growth regulator.

Net assimilation rate (NAR) synonymously called as “unit leaf rate” expresses the rate

of dry weight increased at any instant time on leaf area basis with leaf representing an estimate of the size of the assimilatory surface area. Gregory (1926) was first to suggest the use of this function in the analysis of growth, there on used by several workers in affluent crops. In general, application of growth regulators showed the higher value for NAR at all the growth stages. GA3 @ 20 ppm significantly increased the NAR, this may be attributed to increase in the dry matter production by maintaining more number of green leaves. Growth regulator Triacontanol significantly increased NAR in Ocimum carnorum (Gupta et al., 1995). Baghel and Yadava (1994) reported that foliar application of NAA significantly increased NAR in blackgram.

5.4 Anatomical parameters

Aromatic oils are secreted in internal glands or in hair like structure known as trichomes. In patchouli there are well defined oil glands in which patchouli oil is secreted. The amount of aromatic oil extracted from the leaf of patchouli is positively correlated with the number of oil glands present in the mesophyll cells of leaf. However, Singh and Hippalgaonkar (1993) have stated that it was difficult to calculate leaf oil gland number precisely and further opined glands did not appear to account for the differences in essential oil yield. On the contrary, in the present investigation there was a positive correlation between leaf oil gland and essential oil yield.

Of the different plant growth regulators tested NAA @ 20 ppm and GA3 @ 20 ppm

was found most effective in increasing oil gland number at 90 and 120 DAP. This might be due to increased leaf area and leaf area index due to application of PGRs as observed in the results pertaining to growth parameters. The increase in oil gland number by growth regulators has been attributed to be due to increase in length of palisade cells and width of epidermal cells in jasmine (Pappaiah and Muthuswamy, 1980).

Similar results were obtained in patchouli by Singh and Hippalgaonkar (1993) who

recorded increase in gland number by application of different concentration of TIBA over control.

5.5 Biochemical parameters

Crop yield is mainly dependent on the interplay of various physiological and biochemical functions of the plant in addition to the impact of growing environment. The cause and effect relationship is difficult to understand mainly because of complexity in understanding the interplay of several processes and functions which ultimately lead to changes not only in growth, development and physiology. It was observed in the present study that the treatments differed significantly with respect to total chlorophyll content, phenols content, nitrate reductase activity whereas carotenoid content failed to show any significant difference among treatment in any of growth stage.

The effect of growth regulators on total chlorophyll content in leaf exhibited significant

differences only at 90 DAP. The application of GA3 @ 20 ppm and GA3 @ 40 ppm resulted in significantly higher chlorophyll content at 90 DAP. Increased chlorophyll content of leaves of GA3 treated plants may be indication of increased rate of photosynthesis (Kanjilal and Singh 1998) and decreased chlorophyll degradation and increased chlorophyll synthesis and the increase in total chlorophyll content can also be attributed to involvement of growth regulators

in promoting the synthesis of chlorophyll as well as development of chloroplast (Fetcher and Cullagh, 1971).

These results were in accordance with Kanjilal and Singh (1998) who explained that

application gibberlic acid increased chlorophyll content in chamomile. Similarly Singh and Hippalgaonkar (1993) explained application of kinetin increased total chlorophyll (a+b) content.

The carotenoid content in leaves (mg/100 g fresh weight) did not exhibit significant

difference among the treatments. On an average, a total carotenoid content of 10.73, 13.62, and 13.68 was obtained at 60, 90 and 120 DAP respectively. However, Kanjilal and Singh (1998) reported that on an average 10.36 in chamomile.

Significant differences were observed among the treatments with respect to phenol

content at 90 and 120 DAP wherein NAA @ 20 ppm recorded maximum phenol content at 90 and 120 DAP. This increase in phenol content at later stages plant growth may be due to enhancement of synthesis of phenolic glycosides by application of chemicals to yield free phenols (Sharma et al., 1983). Similar observations were recoded by Chakraborthy et al. (2002).

The enzyme nitrate reductase catalyses the reduction of nitrate to nitrite which is first

step in the assimilation of nitrate by plants. The reaction takes place in the cytoplasm of the cell in both roots and shoots (Kumar et al., 1989).

It has been reported by Bowerman and Godman (1971) that the total dry matter

accumulation is significantly and positively associated with corresponding nitrate reductase activity in clocium. In general NRA was highest during 60 DAP and it declined in later stages. It was observed that activity of nitrate reductase increased significantly in the treatment that received GA3 @ 20 ppm, NAA @ 20 ppm and GA3 @ 40 ppm. The increased nitrate reductase activity may be related to enhanced NR protein and NR-mRNA contents by application of NAA (Christophe et al., 1997). Similar observations were recorded by Charlotte et al. (1982) who reported increased nitrate reductase activity by GA, kinetin and zeatin treatments over control in dwarf bean.

5.6 Yield and yield parameters

Improvement in yield, according to Humphrieg (1979) could happen in two ways i.e. by adopting the exacting varieties to grow better in their environment or by altering the relative production of different plant parts so as to increase the yield of economically important parts. The growth regulators are capable of redistribution of dry matter in the plant, there by bringing about an improvement in yield potential.

The effect of growth regulators on total herbage yield (fresh and shade dried)

exhibited significant differences at harvest. The application of GA3 @ 20 ppm and NAA @ 40 ppm and GA3 @ 40 ppm which were on par with each other in present investigation registered significantly higher yield as compared to control. The higher herbage yield by plant growth regulators may be due to enhancement in yield contributing factors viz., plant height, number of leaves, number of branches and leaf area. GA3 is known to modify growth habits of plants such as elongation of internodes by increasing in size and number of cells, increase in surface size and shape of leaves and elongation of lateral buds (Gonzalex and Gjerstad, 1960) while NAA develops stronger root system causing over loading to transport system.

GA3 and NAA may cause profused foliage and erectrophyl canopy causing more

photosynthate formation. The results were in accordance with Misra (1995) who reported maximum leaf fresh weight in patchouli treated with GA3 @ 25 mg/l. Similarly, Kewalanad et

al. (1998) also reported maximum herbage yield in Japanese mint plant treated with 30 and 40 mg/l of GA3. Mousa and Emary (1983) reported that GA3 at 50 ppm significantly increased fresh herbage in sweet basil. All concentrations of GA3 sprays (100, 200, 400 ppm) significantly increased herb fresh weight of geranium (Mohamed et al., 1983).

Oil per cent recorded significant difference among treatments at harvest where in NAA @ 20 ppm recorded maximum oil per cent followed by GA3 @ 20 ppm. This increase in oil per cent may be related to increased number of oil glands/mm² leaf area. Oil content (kg/ha) recorded significant differences among the growth regulators at harvest where in GA3 @ 20 ppm and NAA @ 20 ppm resulted in higher oil content which were on par with each other. Existence of positive relationship between yield and concentration of essential oil was reported by Rajeshwara et al. (1993).

Increase in biomass and essential oil yields of many aromatic crops through

application of growth regulators were reported by Mohammed et al. (1983) and Singh et al. (1989) in geranium. In the present investigation the application of PGRs resulted in more number of leaves and higher leaf yield which ultimately recorded higher essential oil yield. A direct effect of plant growth regulators on monoterpene metabolism through increased activity of enzyme that synthesize essential oil terpenes leading to accumulation of essential oils was observed in several species belonging to family Lamiaceae. Such a probability existence in the food crops such studies explain higher content of essential oil in aromatic crops also is obtained in the treated plants.

Similar observation were also made by Bhaskar et al. (1997) in patchouli reporting

maximum oil content by application of growth regulator TIBA. Bhattacharya et al. (1995) also reported increase in essential oil content of rose-scented geranium by application of GA3 and NAA and Krishnamoorthy and Madalageri (2000) reported increase in essential oil content in ajowan by NAA application.

All these studies clearly explain application of growth regulators influence the growth

and yield irrespective of type of crops. Thus, an increase in the essential oil content in patchouli due to PGRs is not an exception.

Future line of work

Based on results obtained the present investigation the following suggestions have indicated for further studies.

1. It is necessary to identify suitable growth regulators and its concentration for improving

herbage production and oil content (%) in patchouli 2. It is also important to study the interactions between externally applied growth regulators

and endogenous hormones in manipulation of source sink relationship 3. It is also important to study the combined effect of growth regulators and micronutrients

spray on growth, physiology and yield of patchouli

VI. SUMMARY

A field experiment was conducted at Main Agricultural Research Station, University of Agricultural Sciences, Dharwad during kharif 2004 to study influence of different growth regulators on various morphological, biochemical anatomical and yield and yield components in patchouli. The experiment was laid out in randomized block design (RBD) with three replications. The salient findings of the investigation are summarised here under. 1. The plant height increased significantly due to NAA @ 20 ppm followed by GA3 @ 20

ppm as compared to control. The treatment mepiquat chloride @ 500 ppm and mepiquat chloride @ 1000 ppm reduced the plant height.

2. Number of branches and number of leaves were increased significantly with GA3 @ 20 ppm and NAA @ 20 ppm showing profound effect of growth regulators.

3. Leaf dry matter was maximum during harvest. The application of growth promoters GA3 @ 20 ppm, NAA @ 20ppm increased leaf dry weight significantly when compared to control.

4. All the treatments showed significant increase in stem dry weight and maximum was noticed at harvest. Among various treatments tried, maximum stem dry weight was noticed in GA3 @ 20 ppm and NAA @ 20 ppm.

5. Total dry matter increased significantly due to application of growth regulators. The treatments with GA3 @ 20 ppm and NAA @ 20ppm recorded significantly higher values for TDM over all other treatments at all stages except 60 DAP.

6. The significant increase in leaf area, leaf area index, leaf area duration was noticed in all the treatments at all the stages except 60 DAP over control. GA3 @ 20 ppm recorded highest LA, LAI and LAD.

7. Specific leaf area, specific leaf weight differed significantly at 90 DAP among treatments. Among the treatments GA3 @ 20 ppm and NAA @ 20 ppm recorded higher SLA and SLW over other treatments and control.

8. Application of growth regulators GA3 @ 20 ppm significantly increased AGR, CGR, NAR at 60-90, 90-120 and 120 DAP to harvest whereas RGR was significantly increased with GA3 @ 20 ppm at 60 –90 and 90-120 DAP.

9. A significantly higher number of oil glands was observed at 90 and 120 DAP. NAA @ 20 ppm exhibited highest number of oil glands.

10. Total chlorophyll content differed due to growth regulators. Total chlorophyll content were significantly higher with GA3 @ 20 ppm and GA3 @ 40 ppm at 90 DAP.

11. The results on carotenoid content by application of growth regulators were non-significant.

12. The significant difference among the treatments with respect to phenol content was noticed at 90 and 120 DAP. NAA @ 20 ppm recorded higher phenol content over other treatments.

13. Nitrate reductase activity was maximum at 60 DAP and decreased thereafter. The application of growth promoters GA3 @ 20 ppm and NAA @ 40 ppm increased NRA significantly @ 90 and 120 DAP when compared to control.

14. The results on various yield and yield attributes indicated that all the yield contributing characters viz., fresh and shade dried herbage yield, essential oil per cent and essential oil content (kg/ha) increased significantly due to growth regulator treatments GA3 @ 20 ppm and NAA @ 20 ppm.

15. Based on the above results, it is concluded that, the application of GA3 @ 20 ppm and NAA @ 20 ppm was more effective in increasing the yield potential.

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EFFECT OF PLANT GROWTH REGULATORS ON GROWTH AND YIELD OF PATCHOULI (Pogostemon

cablin Benth. L.)

ANILKUMAR M. 2005 Dr. A. S. NALINI PRABHAKAR MAJOR ADVISOR

ABSTRACT

A field experiment was conducted at Main Agricultural Research Station, University of Agricultural Sciences, Dharwad during kharif 2004 to study the influence of different growth regulators, on various morphological, physiological, biochemical, anatomical, yield and yield components in patchouli. The experiment was laid out on randomized block design using four growth regulators (GA3, NAA, Mepiquat chloride and Miraculan) at different concentrations with three replications.

Growth regulators significantly influenced the morphological characters such as plant height, number of branches per plant and number of leaves per plant. Further number of leaves and number of branches per plant was significantly higher with GA3 (20 ppm) followed by NAA (20 ppm). The leaf dry weight and stem dry weight were significantly higher with application at GA3 (20 ppm) followed by NAA (20 ppm).

The growth regulator GA3 (20 ppm) and NAA (20 ppm) revealed significantly

increased LAI, LAD, AGR, CGR and NAR at 90, 120 DAP and at harvest. The results on various biochemical parameters on anatomical parameters revealed

significant differences among treatments at 90 and 120 DAP except carotenoid content. Among growth regulators GA3 (20 ppm) followed by NAA (20 ppm) recorded higher chlorophyll content. Higher NRA, higher phenol content and higher number of soil glands.

The results of various yield and yield attributes indicated that all the yield contributing

characters viz., fresh and shade dried herbage yield, essential oil per cent and essential oil content (kg/ha) increased significantly due to growth regulator treatments. It is concluded that, the application of GA3 (20 ppm) followed by NAA (20 ppm) was more effective in increasing the yield potential.

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