CHAPTER – IV

METHODS AND MATERIALS

The present investigation entitled, “To Study the Effect of Inorganic and
Organic Sources of Nutrients on Yield and Quality of Lemon Grass
(Cymbopogon flexuosus L.) ” involved field experimentation conducted for two
consecutive years at college Agricultural Research farm Nidharia in Ballia district
(U.P.). The details of material used and methods employed during the course of
investigation are mentioned in this chapter.

4.1 Soil:

4.1.1 Experimental sites:

The micro survey of the area under study was undertaken and sampling
sites were demarcated. Sampling sites were carefully chosen taking into
consideration the ground water, micro-relief, degree of erosion, surface drainage,
proximity of trees and all other factors likely to affect the soil in comparison to
normal type. Field experiments were conducted at the Agricultural Research farm
Nidharia of S. M. M. Town Post Graduate College Ballia during 2007-2008 and
2008-2009.

4.1.2 Soil sampling:

Soil profiles were exposed with the help of spades. At each location
composite soil samples were obtained from different horizons, viz. 0-15, 15-30,
30-45 , 45-60 and 60-90 cm segments. Collected composite soil samples from each
depth, crushed up into smaller pieces and mixed together and the small amount of
soil sample of about 1 kg which was representative of the original material
(quartering method) was taken. The samples were placed in numbered calico bag
giving details of location and depth of soils. The samples were brought to
laboratory for analyses work.

Soil samples from experimental plots were also taken before transplanting
(One month after addition of amendments) and at every harvesting stage.

4.1.3 Preparation of soil samples in the laboratory:

The large lumps were broken and spread it on a water proof sheet to make
it air dry and passed through a sieve with round holes of < 2mm diameter, mixed,
weighed and stored in plastic bag. Placed a label inside the plastic bag separately
for analyses.

4.1.4 Analyses of soil samples:

Physicochemical analysis of soil like pH, ECe, organic carbon, cation

exchange capacity (CEC), calcium carbonate, N, P, K, were performed as per
standard methods mentioned as follows:

4.1.5 pH

A soil water suspension was prepared in the ratio of 1:2.5 (10 gms of soil
and 25 ml distilled water) and pH was recorded with the help of glass electrode
digital pH meter, (Chopra and Kunwar 1982).
4.1.6 Electrical Conductivity (E.C.)

A soil water suspension was prepared in the ratio of 1:2.5 (10 gms of soil
and 25 ml of distilled water) and EC was measured with the help of Conductivity
Bridge at room temperature. The scale reading (mhos cm-1) was converted to dSm1
using the following formula, (Chopra and Kanwar 1982):

EC (d Sm-1) = Scale reading x 1000 x cell constant

4.1.7 Organic Carbon

Soil organic carbon content was determined by Rapid Titration Method

(Walkley and Black, 1934). In this method two grams of soil was oxidized with a
mixture of potassium dichromate and concentrated sulphuric acid utilizing the heat
of dilution of sulphuric acid. Unused potassium dichromate was back-titrated with
ferrous ammonium sulphate in presence of diphenylamine indicator.

4.1.8 Cation Exchange Capacity (C mol (p+) kg-1)

Cation exchange capacity was determined using sodium acetate solution

(pH 8.2) with the help of centrifuge, (Jackson 1973).

4.1.9 Available Nitrogen

Available nitrogen was determined using alkaline potassium permanganate

method (Subbiah and Asija, 1956). The procedure involves distilling the soil with
alkaline potassium permanganate solution and determining the ammonia liberated
by titrating against sulphuric acid (0.02N).

4.1.10 Available Phosphorus

 The Olsen’s method (Olsen et al., 1954) was used for the determination of
available-P in soil. In this method soil was extracted with 0.5 M NaHCO3 (pH 8.5).
Five ml of extract was taken and colour was developed by ascorbic acid solution.
After 10 minutes, the intensity of blue colour was measured on spectrophotometer
at 720 nm.

4.1.11 Available Potassium

Five gms soil was extracted with 1N neutral (pH 7.0) ammonium acetate
solution by shaking for 30 minutes potassium content in the extract was
determined flamephotometrically (Jackson, 1973).

4.1.12 Standard methods

The following standard methods were adopted for the analyses of soil and
plant as per the objectives and sampling –

Parameter Method Reference

Soil analysis-
pH 1: 2.5 ratio of soils & water M.L. Jackson (1973)
EC(dSm-1) 1: 2.5 ratio of soil & water M.L. Jackson (1973)
Organic C (%) Wet digestion method Walkley &Black’s (1934)
CEC( cmol(+p)/kg NH4OAc method M.L. Jackson (1973)
Available N (kg/ha) Alkaline KMnO4 method Subbiah & Asija (1954)
Available P (kg/ha) 0.5M NaHCO3 extractable Olsen’s et al (1959)
Available K (kg/ha) NH4 oAc extractable Muhr et al (1973)
Available S (kg/ha) Turbidity by BaCl2 Chesnin &Yen (1959)
Ca++ (g/L) Titration method Versenate (EDTA) method
Ca++ + Mg++ (g/L) Titration method Versenate (EDTA) method
CO3-- and HCO3- Titration method M.L. Jackson (1973)
(g/L)
 Cl- (g/L) Titration method M.L. Jackson (1973)
SO4-- (mg/kg) Turbidity by BaCl2 Chesnin &Yen (1959)

4.2 Observations Related to Aromatic Crops:

4.2.1 Experimental site : Agricultural Research farm Nidharia (site
A)Block – Hanumangang,
District – Ballia.

4.2.2. Soil : Sandy Lome

4.2.3. Soil Texture : Clay loam

4.2.4. Crops : Lemongrass(Cymbopogonflexuosus)

4.2.5. Year : 2007-2009

4.2.6. LAY OUT OF EXPERIMENT

The experiment was conducted during summer, Kharif seasons of 2007-

2008 and 2008-2009. The details of lay out (Fig. 1) are as follows :

4.2.7 Details of Layout

Experimental design : R.B.D.
Number of treatment combinations : 15 (5x 3)
Main plot treatments : 5
Sub plot treatments : 3
Number of replications : 3
Total number of plots : 15
Plot size : 5 x 3 m2
Width of replication border : 1.0 m
Width of main irrigation channel : 1.0 m
Width of sub-irrigation channel : 0.5 m
Test crop : Lemon grass
Spacing : 60 x 60 cm2

4.2.8. Details of treatments:

Treatment combination –

T1 – 100% NPK as RDF (40:60:40 Kg ha-1)

T2 – 50% NPK+ 5t FYM/ ha-1.

T3 – GLM (Green leaf manuarying) 10t / ha-1.+10t FYM

T4 – 100% N through vermi compost ha-1

T5 – 100% N through poultry manure ha-1

4.2.9. Layout:

The details of the layout plans are given in Fig 1.

Lay out

T1 T5 T2

T5 T3 T4

T3 T1 T3

T2 T4 T5
 T4 T1 T2
3x3m

v1 V2 V3

Experemental plot

T1V1

4.2.10. Preparation of fields:

At proper moisture condition the land was ploughed once with tractor
drawn mould board plough followed by two disc ploughings. Each ploughing was
followed by blanking stoves. Weeds were removed by conventional methods and
operations were carried out to prepare the land uniformly for the field trial.

4.2.11.Application of organic amendments:

FYM , Green leaf manure, Vermicompost and poultry manure was applied
at the time of field preparation at per recommendation of treatment.

Table -Physical and chemical properties of the soil of experimental site

Sl.No. Particulars Value Method adopted and reference

I. Physical properties
1. Coarse sand (%) 28.94 Hydrometer method
2. Fine sand (%) 8.54 (Piper,1966)
3. Silt (%) 44.67
4. Clay (%) 17.85
II. Chemical properties
1. Soil pH 8.7 pH meter (Piper,1966)
2. Electrical conductivity (dS/m-1) 7.1 ConductivityBridge (Jackson,1967)
3. Organic carbon (g/kg) 1.9 Wet oxidation method (Jackson,1967)
4. Available nitrogen (kg/ha) 235 Alkaline permanganate method
(Subbaiah and Asija, 1956)
5. Available phosphorus (P2O5 18.5 Olsen’s method (Jackson,1967)
Kg/ha)
6. Available potassium (K2O 227 Flame photometer
Kg/ha) (Jackson,1967)
III. Soil moisture
1. Initial soil moisture (%) 18.67 Gravimetric method

4.2.12. Preparation of root slips and Propagation of plants:

The crops were propagated by means of rooted slips. Which was obtained
from well-grown clumps. Tops of clumps were cut off within 25 cm of the root.
The lower brown sheath was removed to expose young roots. and the old roots
were clipped off keeping the slip 25-30 cm long. The culm (root portion) was
divided into slips containing 2-3 tillers. Two slips were placed into each hole about
15 cm deep. Rooted slip of lemon grass was planted at a distance of 60 cm. in row
to row and 60 cm. plant to plant. The plantation was done on July 31, 2008. Field
was irrigated immediately after planting.

4.2.13. Application of fertilizers:

Recommended Doses of Fertilizer 20 kg nitrogen through urea, 60 kg

phosphorous through single super phosphate (SSP) and 40 kg potassium through
muriate of potash per hectare as basal dose at the time of plantation. Remaining
nitrogen (20 kg) was applied as top dressing in 3 split doses during the each
growing season.

4.2.14. Irrigation and weeding:

 The plots were flood irrigated in 15 to 20 days during dry period and 22 to
28 days during rainy season as required. A protective irrigation could be made in
February- June. After every irrigation, weeding was done with the help of spade to
maintain proper fertility level of the soil and removal of weeds from the field.
Irrigation is given immediately after planting when planting is done in dry days.
Thereafter two irrigations are given at 10 days interval to establish the crop.

Pest control and care - As per requirement for the pest infestation Methodology
for analysis of soil and plant –

4.3 Observations:

4.3.1 Plant Height :

The height of plants in all the plots was measured from ground level to the
tips of tallest leaf at harvesting stage and mean height per plant was recorded. The
plant height of aromatic crops were measured at 30, 60, 90, 120 and 150 days after
transplanting; 30, 60 and 100 days after first harvesting; 30, 60, 90 and 120 days
after second harvesting; 30, 60 and 120 days after third harvesting and 30, 60 and
90 days after fourth harvesting and tillers were also counted on same dates.

4.3.2 Number of Effective Tillers

The number of effective tillers per square meter was counted in each plot.

4.3.4 Harvesting:

The first harvesting was done at 150 days after transplanting (DAT).
Subsequent harvestings were done at interval of 120 days after first harvesting, 100
days after second harvesting, 100 days after third harvesting and 90 days after
fourth harvesting. Harvesting was done with the help of sickles. The plants were
cut close to their bases about 10 cm above ground level.

4.3.5 Plant sampling:

Plant samples were also taken at all the harvesting stage for the
determination of mineral composition.

4.3.6 Chemical analysis of plant samples:

Air dried plant samples were ground in Wiley Mill having stainless steel
blades and digested in diacid mixture of H2SO4 : HClO4 (10:4) as per standard
procedures (Jackson, 1967) were followed for the estimation of phosphorous
(colorimetric), potassium (flam photometeric), Sulphur (Spectro photometer),
Carbonate and bi-Carbonate (Titration method) and Nitrogen by micro-Kjeldahl
procedure.

Fig.02 - Ploughing of Experimental Plots.

Fig.03 - Preparation of Vermicompost/FYM.

Fig.04 - Processing of Poultry Manure.

 Fig.05 - Integration of Manures (Urea, DAP, MOP)

Fig. 06 - 30 Days old Lemongrass. Fig.07 – 60 Days old Lemongrass.

Fig. 08 - 120 Days old Lemongrass. Fig.09 – 150 Days old Lemongrass.
 Fig. 10 - One Hundred Eighty Days old Lemongrass.

4.3.7 Steam distillation of lemon grass (Cympopogon Spp) :-

Crude lemon grass oil was obtained by steam distillation of grown

lemongrass Sp. Following the method described by kaul et al 2004. To obtained
the maximum yield of oil and to facilitate release of oil, the grass is chopped into
shorter lengths and is filled in the distillation still and it’s lid is fitted tightly by
swivel bolts, so that oil and vapour do not leak out. The steam is injected in the stil
with the help of steam spargers provided at the bottom of the vessel.

The upcoming steam carries away the oil form the plant materials i,e
lemongrass and both oil as well as steam pass to the condenser through vapour
line, where these vapours get condensed and oil and water are separated in the
separators. Oil being lighter is separated out from the top and water being heavier
is taken out from the bottom of separators. The steam distillation process took
about 2.5 to 3 hours. The oil is further decanted and filtered. The oil was stored in
glass containers. Containers should completely be fitted with oil contains
excluding any air and protecting from sunlight.
4.4 Statistical analyses:

The observed data were analyzed statistically in order to judge the

significance of treatments at 5 per cent level of significance. Values of correlations
co-efficient among different factors were also worked out.

Soil & Climate

It flourishes in a wide variety of soils ranging from rich loams to poor

laterite; also suitable to ‘jhum fallow’, hill slopes and flood free degraded land,
best suited to well drain sandy loam. Water logged conditions should be avoided as
they are unsuitable for its cultivation. It requires a warm humid climate with plenty
of sunshine and rainfall ranging from 1800 to 3000 mm. High temperature and
sunshine are conducive to the development of oil in the plant.

Varieties

The following varieties were studied for the commercial cultivation of

Lemongrass:

(i) For irrigated condition C.K.P-25 released by RRL, Jammu

(ii) Krishna developed by CIMAP both for irrigated and rainfed and also
hill slopes

(iii) Nima a dwarf variety developed by CIMAP found promising. The

citral content of Nima is much higher than Krishna. Oil recovery is in
between 0.8-1.0 %.

(iv) Another suitable variety for NE region is Jor Lab L-2 developed by
RRL, Jorhat.
Planting :

Planting was done in May-June. However, with irrigation planting can be

done during any month of the year except December-January. One or two slips are
planted into each hole, about 5-8 cm deep. Slips were transplanted firmly into the
ground. Plants got established well within 25-30 days after planting.

Manures and fertilizer application:

During dry season after each harvest one irrigation and subsequently
application of recommended dose of fertilizers were to be followed for optimum
herb production.

Spacing :

Generally, a row spacing of 20 cm with a row width of 40 cm, would

required along with a total of 125 000 plants per ha in a high rainfall area or under
irrigation.

Planting depth :

Slips were be planted at a suitable depth to cover the root zone properly
and the soil would be firmed around the plant to remove trapped air pockets.

*******