LabManual2012 PDF

INDEX
S. No. Title Page No.
01 Importance of Soil Testing, Collection of Soil Sample, its 1-4

Processing and Handling in Laboratory
02 Sampling, Processing and Storage of Plant Samples 5-7
03 Simultaneous Measurement of Bulk Density and Water 8-9

Content by Nuclear Methods
04 Estimation of Soil Moisture by Different Methods 10-12
05 Determination of Soil pH and Electrical Conductivity 13-14
06 Determination of Soil Organic Carbon 15
07 Determination of Available (Mineralization) Nitrogen in Soil 16-17

by Alkaline Permanganate Method
08 Determination of Total Nitrogen in Soil and Plant 18-19
09 Determination of Phosphorous in Soil and Plant 20-22
10 Determination of Potassium in Soil and Plant 23-24
11 Determination of Sulphur in Soil and Plant 25-26
12 Determination of Zn, Cu, Fe and Mn in Soils and Plants by 27-30

Atomic Absorption Spectrophotometer
13 Estimation of Boron in Soils, Plants and Water 31-32
14 Profile Studies of Deep Black Soils 33-36
15 Food Preservation by Gamma Irradiation and its Importance 37-42
16 Plant Tissue Culture Techniques for Mass Propagation of 43-46

Banana
17 Estimation of Microbial Biomass Carbon in Soil 47
18 Application of Global Positioning System (GPS) 48
Appendix - I to VI 50-57
 CAFT on Advances in Agro-technologies for Improving Soil, Plant & Atmosphere Systems from 11-31 Oct. 2012

Importance of Soil Testing, Collection of Soil Sample, its Processing and

Handling in Laboratory
P.S. Kulhare
Department of Soil Science and Agricultural Chemistry
College of Agriculture, J.N.K.V.V., Jabalpur
Assessment of a soils fertility status involves (ii) Grouping of soil for their classification
(iii) To determine the specific soil problem
an estimation of its available nutrient status
such as an acidity, alkalinity and sodicity
i.e. the portion or amount of nutrient directly
if exist. Subsequently giving
available in soil for subsequent uptake by
recommendation for their correction
crop plant. This exercise commonly referred
(Lime/Gypsum requirement etc.)
to as soil testing and is used to arrive at
(iv) To predict the probability of getting
optimum fertilizer application ratio. The need
maximum response of crops to
for estimation of available nutrient arises
fertilizers.
because only a small fraction of what the soil
contains is the total nutrient content of the Procedure for soil testing
soil. Soil test are calibrated by correlating
The procedure for testing the soil to
them with crop response and the result from
meet these objectives is divided into the
the basis for making fertilizer
following phases:
recommendations.
(i) Collection of soil samples and its
Estimation of nutrient contents and
preparation
forms in materials that are involved in
(ii) Extraction and determination of nutrients
nutrient supply and dynamics is a conical step
and physico-chemical properties of the
towards planning scientific nutrient
soil.
management. In this content, both soil and
(iii) Interpretation of analytical results.
plant testing information comes out of the
(iv) Recommendation and follow up of
interpretation of analysis assumes a greater
results and evaluation of
value when their concentrations and amounts
recommendations.
can relate to soil fertility, nutrient availability,
plant growth, yield and quality of the crop Soil testing is a chemical method for
produce. estimation of nutrient supplying power of a
soil/ soil fertility evaluation.
Why soil testing ?
Soil fertility may be defined as the
• Soil fertility status assessment involves an capacity of soil to furnish available plant
estimation of its available nutrient status nutrients to the plants in proper amount and
appropriate balance, under ideal condition of
• It gives the amount of nutrient directly plant growth. Whereas, Soil productivity is
available in soil for subsequent uptake by the capacity of soil to produce under specific
crop plant. condition of crop production.
• Guides to arrive at optimum fertilizer
application. Advantages of soil testing :
• It is a method of evaluating nutrient status  More rapid method as compare to
(physico-chemical properties) of the soil biological or deficiency symptoms/ plant
i.e. the assessment of the fertility of the analysis.
soil to determine nutrient deficiencies.  One may determine the need of the soil
before the planting of crop.
• It is also concerned with environmental
 To determine the suitability of the soil
quality for the community hazards.
for laying gardens.
Objectives  Lime problems.
 Soil survey.
(i) To evaluate soil fertility and its
The error in soil sampling in a field is
productivity by the estimation of level of
generally greater than the error in laboratory
nutrient (Low, Medium, High).
JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 1


analysis. The most recommendation call for Collection of representative soil sample :
soil testing of each field is about every three  Depending on field conditions and the
years with more frequent testing on lighter objective of sampling, select proper
soils. Therefore, it is necessary that the soil sampling tool (s).
sample should be representative of the area.  Based on difference in soil type, colour,
Further, the subsequent handling operation in crop growth or slope, divide the area in
the laboratory should be carefully performed different homogenous units.
because a minute quantity (1 – 10g) of the
large soil mass of the field is actually used for
the analysis in the laboratory. Unless one is
sure of representative and proper sampling,
the results obtained in the laboratory analysis
will be of no use under the field conditions.
Apparatus and materials :
 Khurpi DIVISION OF AREA
 Spade
 Augers  In the uniform field, demarket the
 Plastic bowl sampling points in a zig zag fashion or
 Scale randomly in such a way that the whole
 Rack field should be covered i.e. about 30-35
 Wooden roller sample per ha.
 Mortar and pestle
 Sieve
 Polythene/paper/cloth bags
 Labels
 Card board cartons
 Aluminium boxes
SELECTION OF SAMPLING POINTS
 At the sampling site, remove the surface

litter with Khurpi or spade. With the
help of the sampling tool (Auger) collect
a sample in a plastic bowl.
 If the soils is hard, make a ‘V’ shape cut
upto 15 cm depth. Remove the soil of
the pit. Now scrap or remove 1 cm / 1”
soil from the surface upto 15 cm depth
from both the side with the help of
khurpi. This scraped soil is collected in a
Screw Auger plastic bowl. This sample is known as
‘primary’ sample. Such primary samples
should be approximately the same
weight.
Parts of an Auger COLLECTION OF SOIL SAMPLE BY

KHURPI

 After collecting at least 30 – 35 primary When labels are hand written special
samples, mix all the samples in plastic care should be taken to prevent ambiguity.
bowl thoroughly and draw about ½ to 1 For arabic numbers “6” and “9” and
kg composite sample by quartering combination like “69” and “96” should be
method. Label the sample in the bowl under line, and ‘1’ and ‘7’ should be clearly
and divide the sample approximately 4 distinguished. Where letters & numbers are
equal part. Discard the 2 opposite used ‘5’ may be easily confused with ‘S’.
portions of the samples and remaining 2 This is why printed labels are always better.
portions are again thoroughly mixed and Field samplers have their own
again divided in to 4 equal parts and 2 numbering system and may have record book
opposite parts again discarded. This containing printed forms for entry of
procedure is continue until ½ to 1 kg information serially numbered. These ‘sample
sample remain in the bowl. This is number’ are main identification of soil for
known as composite sample which is most purposes but the relevant form should
true representative of the area. also have a record of the ‘bag number’ and
 The most suitable containers for soil subsequent ‘laboratory number’.
samples are polythene bags 6x9’’ made When a box of samples is dispatched
of film about 0.13 mm thick, which may to the laboratory, it should contain a packing
be sealed by twisting or tying the neck or note giving the total number of sample,
by mean of rubber bands or adhesive ‘sample number’ of each sample and its
tape. corresponding ‘bag number’, the depth of soil
 If the soil is to be kept in moist condition sample from profile pit and other information
for moisture determination, bacterial needed by the laboratory staff for registration
count and nitrate estimation etc. air tight purposes, particularly on the analysis
containers are preferred. required. A duplicate packing note should be
If the soil to be used for the estimation of sent separately so that missing boxes can be
micronutrients like Zn, Cu, Fe, Mn. Use of investigated.
metallic tools should be avoided. Use sharp On arrival of the soil samples at the
stick or stainless steel. Brass sieve should be laboratory, the content of a box should be
avoided. Nylon net or aluminum sieve should checked against the packing note if any
be used. discrepancies should be reported to the
In majority of cases, large stones and sampler. The samples are register in
pieces of gravel (7.6 – 2mm) can be laboratory giving each sample a ‘laboratory
discarded. Soil is broken up and spread out in number’ for particular analysis.
a thin layer on strong paper or polythene film, Small laboratory simply the
preferably on a rack of wire mesh to allow air numbered serially as they arrive. Larger
to circulate. The drying area protected from laboratory may have 2 or more numbering
direct sun and wind. system, using a prefixed letter (group of
Soil samples should be labelled/ letter) to distinguish them. This procedure
numbered by field staff with water proof ink helping to channel samples into various
or paint. These bag numbers being entered in analytical stream. Larger laboratory may need
the sampler’s record books, as each sample is to ‘punched card system’ or other means of
taken together with other information. He storing complete information on all samples.
needs to identify and describe the samples. So that can be recovered quickly.
If labels are used, they should either It is essential to keep a record of the
printed numbers on them already or water date of arrival and the source of all samples.
proof ink should be used to write information A table can be drawn up for each month.
on them (this excludes pencil, washable ink Information sheet: The soil sample thus
pens and ball point pens). Duplicate labels collected must be furnished important
should be placed between the two bags, never information like –
in the bag with the soil. The information on a 1. Sample number
label should be kept to a minimum preferably 2. Name and address of the farmers.
a number which may be either a ‘bag number’ 3. Details of the field and site. Local name
or ‘sample number’. Depth may be given of field, Khasra no etc.
usefully for soil profile samples. 4. Date of sampling


5. Name of crop and variety to be sown 5. Keep the sample in a clean bag.
6. Source of irrigation 6. A sample should not be taken from large
7. Whether the crop in the subsequent area (more than 1-2 ha).
season will be irrigated or un-irrigated. 7. Sample for micronutrient analysis must
8. Name of crops and fertilizer used in be collected by steel or rust free
previous years. khurpi/auger and kept in clean polythene
9. Date of harvest of the previous crop. bag.
10. Any other problem observed in the field. 8. Avoid sampling from low – lying spots,
manure dumping sites, near trees and
Preparation of soil sample for testing
from fertilizer placed zones.
1. Spread sample for drying on clean cloth, 9. Use clean bags for sample collection. Do
plastic or brown paper sheet. not use bags which had earlier contained
2. Remove the stone pieces, roots, leaves & fertilizer, manure or plant protection
other un-decomposed organic residues chemicals etc.
from the samples. STORAGE :-
3. Large lumps of moist soils should be  The registered and labelled samples in
broken. laboratory are finally placed in a
4. After air drying the samples should be cardboard carton. Label the carton
crushed gently and sieved through a 2 properly with the details of soil sample
mm sieve. and store in a separate room. The room
5. About 250 g of sieved sample should be should be away from direct sunlight/wind
kept in properly labeled sample bag for or dampness.
testing.  The room exposed to heat or cold or
Appropriate time for soil sampling dampness in not advisable.
An ideal time for soil sampling is just REFERENCES:-

after harvest of the Rabi crops,
 Physical and chemical methods of soil
Precautions to be taken during collection and water analysis FAO soils bulletin
of soil sampling 10. Food and Agricultural Organisation
of United Nations.
1. Remove all debris from surface before  Soil Fertility and Fertilizers: Samuel
collection of soil sample. Tisdale and Warner Nelson, Machmillan
2. Avoid taking sample from upland and Pub. Newyork.
low land areas in the same field.  Analytical Agricultural Chemistry: S.L.
3. Take separate sample from the areas of Chopra and J.S. Kanwar.
different appearances.
4. In row crop take sample in between rows.


Sampling, Processing and Storage of Plant Samples

G.D. Sharma
Sampling Drying
A sample must be a true After washing, the excess water in
representative of the crop under field the sample may be soaked by lacing them
conditions. The best way to collect a between the folds of filter paper sheets.
representative plant sample is to traverse All the samples should dry as rapidly as
the field diagonally and collect the possible so as to reduce chemical and
samples from the deficient and normal biological degradation if any. Sample
plants. These samples should be should be dried in a hot air oven at 700C.
composited separately representing poor
The material to be dried should be loosely
growth and normal growth. Plants which
packed so as to allow the free movement
are infected with disease or attack of
of moisture laiden air. Oven made of
insects, which are under stress due to
stainless steel shelves should be used.
excess water or drought and those which
Plant samples have been dried generally
are heavily coated with dust should not
for 24 to 36 hours. Care should be taken
form a part of the sample. Extraordinary
to insure that the plant sample is not
care must be taken to avoid contamination
bunched together in isolated area of the
resulting from dust, soil, fertilizer and
oven, as large losses of N any occur. Plant
spray residues etc.
samples may also be dried in a microwave
The samples so collected should oven. This procedure is repid and can dry
be transferred to paper bags indicating the individual samples to a brittle state in as
sample number and/or the field number or little as 5 minutes.
other details. In case the sampling site is
Grinding and storage
far away from laboratory and requires
longer travel period, the sample may be The oven dried samples should be
placed in polyethylene bag and group in a grinder, fitted with stainless
transported in an ice chest. Low steel blades so as to pass the samples
temperature will minimize the through a 40- mesh sieve. After grinding,
physiological activity and also check the the plant sample should be mixed
spoilage of sample due to higher thoroughlyand transferred to polyethylene
temperature and high humidity. or paper bags, labeled clearly and then
stored in the room meant for plant
Processing and storage of plant samples samples. Avoid regular type of grinder as
Washing it normally provided contamination. The
The fresh plant samples brought to sample grinder consisting of hard plastic
the laboratory should be immediately can be used for grinding (such type of
washed in order to make them free from grinder is developed by tecator).
dust or any other adhering substance. Precautions
Samples should first be washed under the
1. Avoid sampling the plants which are
running tap water and if necessary.
infected with disease and/or insects
Subsequently, these samples should be
or those suffering from effects of
washed with acidified distilled water (1
excess or scarcity of water.
ml concentrated HCl/litre) followed by
thorough rinsing twice with deionized 2. Before subjecting the samples for
water or distilled water. testing, decontaminate the collected


samples by thorough washing, and The plant sample is diacid with

rinsing. mineral acid mixture (conc. HNO3, &
3. Washed samples should be dried as HClO4) and heated for more rapid
rapidly as possible. The samples decomposition. The volatile constituents
should not be packed tightly during disappear and non-volatile mineral
drying in an oven and the elements enter into solution.
temperature should not exceed 700C. The heating is continued until
4. The grinder to be used for grinding digest is reduced to few ml of clear white
the samples should have stainless residue. The residue is dissolved in HCl.
steel blaces or hard plastic to avoid The test solution is prepared by dilution
contamination and the ground and filtration.
samples must be passed through a
Reagents
40- mesh sieve to ensure uniform
fineness. For iron determination, It  Diacid mixture. Mix 100 ml of
is advisable to grind the sample by conc. HNO3 and 40 ml of 60%
hand in an agate or porcelain mortar HClO4 or else 30 ml of 72%
or in a mill made of non ferrous HClO4.
alloy if hard plastic grinder is not Procedure
available.
5. The processed samples should be Digestion with diacid mixture: Add 5 ml
kept in polyethylene or paper bags of diacid mixture into the beaker in which
and stored in room free of dust soil 1.0 g plant sample has been transferred in
and smoke etc. the beaker and place on a hot plate for
brisk heating. Keep a portion of the
6. Care should be taken not to rub the beaker mouth open to permit the gases to
plant tissue acid mixured like any of escape. Continue heating until evolution
the washing the samples. of copious dense white fumes subsides
Methods of ashing of plant tissues leaving about 3 ml of colourless solution
in the beaker which on cooling gives a
Wet ashing is accomplished by
whitish residue. If the residue is not white
digestion with HNO3, HClO4 and H2SO4
and signs of beginning of charring are
or else with HNO3 and HClO4 especially
seen, remove the beaker from the hot
for S determination. The method is
plate and let is cool. Then add 2 ml of
unsuited for B determination as the
diacid mixture and place the beaker again
element is volatilized during wet ashing.
on hot plate. Continue heating until the
Dry ashing is done in muffle contents is reduced to about 2-3 ml of
furnace at temperature varying form 400 colourless clear solution. In order to avoid
to 5000C for 2-8 hours. The plant sample excessive heating, a layer of sand may be
taken in platinum or vitreous silica spread over the hot plate. If white residue
crucible is treated with Mg (NO3)2 or is not obtained, the residue is treated
Na2CO3 prior to placing in a muffle again with 2ml of the diacid mixture and
furnace. Dry ashing at 5000C is the heating be continued till the contents
considered satisfactory for the is reduced to about 2-3ml. thereafter, the
determination of Fe, Mn, Cu in biological beaker be removed from the hot plate or
materials. The use of stainless steel lined sand bath. Allow the beaker to cool.
muffle furnaces eliminate contamination
Preparation of plant test solution: Add
from Al and Zn if any during ashing.
5ml of glass distilled water into the
Wet digestion of plant sample with beaker containing digested residue. Heat
diacid mixture the contents gently on a hot plate until
white fumes appear. Remove the beaker


and let it cool. Then, add 10ml of glass If Ca and Mg is to be determined,

distilled water and heat the contents to add 5ml of conc. HCl instead of glass
dissolve the residue. distilled water into the beaker containing
the plant digested residue. Swirl the
Take a 100ml clean volumetric
contents and heat it to warm. Take a clean
flask fitted with a funnel lined with
100 ml volumetric flask fitted with a
Whatman No. 1 paper. Transfer the
funnel lined with Whatman No. 1 filter
contents of the beaker into the funnel
paper. Transfer the contents of the beaker
using a glass rod collecting the filtrate in
into the funnel and collect the filtrate in
the volumetric flask. Rinse the beaker
the volumetric flask. Then, add another
with about 15ml portions of glass distilled
5ml of conc. HCl into the beaker to
water and transfer each rinsing into the
dissolve the remaining residue in the
funnel in order to transfer the digested
beaker. Transfer the solution into the
residue quantitatively. Collect the filtrate
same funnel, collecting the filtrate in the
from each washing into the same
same volumetric flask. Repeat subsequent
volumetric flask. Then wash the residue
washing of the contents of the beaker with
on filter paper with small portions of glass
small portions of 6N HCl and transfer the
distilled water and collect the washing
washing into the same funnel until the
until the volume of filtrate reaches 100ml
filtrate reaches 100ml mark. Similarly run
mark. Stopper the flask, label it and then
a blank digestion and then prepare test
preserve it for the elemental analysis other
solution using conc. And 6N HCl instead
than Ca, Mg and S. similarly run a blank
of glass distilled water as described
digestion and subsequently prepare the
above.
blank test solution following the above
mentioned procedure.


Simultaneous Measurement of Bulk Density and Water Content by

Nuclear Methods
G.P. Tembe
Recent advances is nuclear technology placed between a cesium source and

which have evolved methods for the non detector and then between americium to
destructive determination of the soil water using a combined source but neither
content. Two methods have been used Soanenor other workers (Gardner and
with success. One the neutran scattering Calissendoff, 1967; Gardner, Campbell
method and the other is the y-radiation and Calissendaff, 1969) have attempted to
method. combine the sources in a single collimator
Neutron method : In this method because the higher energy gamma
measurement is made of the number of photons produce Compton scatter through
hydrogen nuclear that are present per unit interaction with the sample and some of
volume of soil and therefore, water this Compton scatter will be counted as
content by volume θ, is measured. gamma rays from the lower energy
Neutrons are uncharged particles having source. The error can be eliminated by
almost the same mass as that of protons or with equipment and method evolved by
that of hydrogen nuclei. A radium- Corey, Peterson and Wakat, 1971). This
beryllium mixture in form of pellets is method is feasible for measuring the
used as source of fast neutrons. The water content and soil density of soil
measurement of soil water by the neutron columns simultaneously when two
method is essentially a measure of the sources are combined in a single
density of the slowed neutron cloud collimator. Measurement of attenuation of
137
developing when Ra-Ba source is inserted cesium and 241americium is done in this
in the soil, the density of the slowed method. Radiation intensity (counts/min)
neutrons being a measure of the soil water after passage through the soil, container
content by volume θ. and container alone is calculated by the
equation (Corey et al.) given in method.
Simultaneous determination of bulk To obtain values of intensity the
density and water content : equipment is required as shown in Fig. 1.
241
With the increased interest in the Am emits a large number of
behaviour of water in swelling soils and gamma rays of various energies but the
resulting theoretical studies (Smile and major energy is 59.6 KeV 241Am has a
Rosenthal, 1968; Philip, 1969) on the half life of 458 years eliminating the need
subject, a method was required for for decay corrections 137CS decays with a
measuring changes in both the water gamma ray of 662 KeV and has a half life
content and soil density in order to of 30 years.
experimentally evaluate these theoretical Detector and pulse height
analyses. Numerous authors have analyzers are used in this system.
proposed measuring the attenuation of Correction for comption scatter
gamma rays of two different energies to Two methods were compared and
non destructively determine both water dual source method evaluated. Known
content and soil density in the same water content and soil densities were
sample. Soane (1967) illustrated the measured. The two soils were Houston
effectiveness of the dual gamma method black, a soil containing monitorillonitic
for measuring bulk density and water clay and cecil a soil containing kaolinitic
content of three soils. The samples were clay. The soil containing different amount


of water was packed to varying densities Neither the soil density nor lengths of
into plastic boxes 7.5 x 4.46 x 4.95 cm. Cecil soil column changed. The column
These boxes were placed between sources of Hoston black soil increased 2 cm in
and detector with long axis parallel to length and the soil density decreased in
beam. Comparisons between the known, top 4 cm.
soil density. The combined source method is
Water content and values applicable to the study of swelling soils,
determined by two methods for four cecil the phenomena of freezing and thawing
soil samples and five Housten Black soil and measurement of water content and
samples are given in table 1. density of the soil inside core barrels.
Table 1: Water content and values References :
determined by twi methods.
Cerey, J.C., Peterson, S.F. and Wakat,
Water content g/cm3 Soil density g/cm3 M.A. (1971). Measurement of
Known Calculated Known Calculated attenuation of 137CS and 241Am
Cecil soil gamma rays for soil density and
water content determinations. Soil
0.001 0.000 0.00 1.831 1.830 1.830
Sci. Soc. Am. Proc. Vol. 35 : 215-
0.044 0.081 0.073 1.712 1.711 1.720 19.
0.120 0.124 0.143 1.999 2.001 1.980 Gardner, W.H. and Callissendortt, C.
0.175 0.118 0.151 2.050 2.067 2.081 (1967). Gamma rays and neutron
Houston Black Soil
attenuation in measurement of soil
bulk density and water content. Pp
0.010 0.000 0.000 1.504 1.504 1.504
101-113. In Isotope and radiation
0.141 0.147 0.139 1.297 1.281 1.290 techniques in soil physics and
0.270 0.284 0.281 1.343 1.367 1.371 irrigation studies. International
0.438 0.472 0.446 1.369 1.349 1.378 atomic agency, Vienna.
Gaedner, W.H., Campbell, G.S. and
0.495 0.532 0.514 1.297 1.234 1.255
Calissendorff, C. (1969). Water
There is little difference between content and soil bulk density
the results obtained by the two correction measured concurrent using two
techniques and method best suited will be gamma photon energies US AEC
determined by the instrumentation Report, RLO 1543-6 Washington
available. State University. Pullman Wash
Following satisfactory evaluation 42 p.
of the dual source method for determining Philip, J.R. (1969). Moisture equilibrium
the known water content and soil density in the vertical in swelling soil. I
of soils, the method was used to Basic theory. Anst. J. Soil Res. 7 :
determine water content and soil densities 99-120.
of Houstan Black and Cecil soil columns Smiles, D.E. and Rosenthal, M.J. (1968).
following infiltration. More water was The movement of water in
added to Houston soil because its greater swelling material. Aust. J. Soil
water holding capacity. Twenty four Res. 6 : 237-248.
hours following the addition of water the Soane, B.D. (1967). Dual energy gamma
transmission measurements were repeated ray transmission for coincident
and the water content and density was measurement of water content and
calculated water remained ponded on dry bulk density of soil. Nature
Houston Black soil at the end of 24 hour 214 : 1273-1274.
period but had infiltrated completely with
the Cecil soil. The two soils responded
quite differently to the addition of water.


Estimation of Soil Moisture by Different Methods

G.P. Tembe
Direct method (Gravimetric method) : Equipments and reagents :

This is the simplest and most (1) Moisture cans
widely used method for measuring soil (2) Glass rod
moisture. (3) Ethyl, methyl or propyl alcohol
(4) Match box
Equipments :
(5) Balance with weights
Sampling tube/auger (6) measuring cylinder
Moisture cans (numbered) (7) Desicator.
Balance with weights/automatic Procedure :
Drying oven
Desicator Weigh about 20-25 gm of wet soil
in a moisture can add about 5ml alcohol
Procedure :
or spirit drop to fully saturate the soil and
Collect soil samples by tube or ignite. Cool the cans in a desicator and
auger from a number of points with in the record the weight. Repeat the process
experimental site and mix thoroughly. several times till a constant weight of the
Place composite sub samples of about 50 ignited soil is recorded. Compute the
gm to 100 gm in soil moisture cans with percentage moisture on the basis of the
tight fitting lids. Take atleast three sub- constant dry weight of the burnt soil.
samples. The moist samples are weighed This method is rapid, reproducible
immediately, dried to constant weight in and more suited for field soil moisture
an oven at 105°C (for about 24 hrs) and determinations where laboratory facilities
reweighed after cooling in a desicator. are not adequate.
Determine the tare weight of moisture Burning of the soil results in loss
cans. Calculate the soil moisture content of organic matter. This method needs to
by determining the loss in weight on be calibrated against oven dry method for
drying and the weight of the oven dry soil each soil type.
as follows : In direct method (By use of neutron
Soil (wt. of wet soil + tare) moisture probe) :
moisture - (wt. of dry soil + tare)
= Rather than taking a sample of
content by (wt. of dry soil + tare)
weight (%) - (tare) soil, it is often desired to measure the soil
moisture status in situ without disturbing
Loss of wt. on drying the system. The neutron moisture meter is
Mw (%) = X 100
Wt. of oven dry soil such a device which is much used in the
field for measuring water content
Percentage of moisture on volume basis
(Belcher, 1952, Gardner and Kirkhana,
= Percentage of moisture on weight basis
1952, Holmes, 1956 and Van Bavel et al.,
x bulk density.
1956). The neutron method is a measure
Gravimetry with drying by burning of the number of hydrogen nuclei that are
Alcohol or spirit (Bouyoucos, 1937) : present per unit volume of soil. Therefore,
In this method, the soil moisture is it is a means of directly observing the
evaporated by igniting the soil mass with moisture content by volume.
alcohol or spirit. The process is repeated
to get constant weight of ignited soil.

Principle : neutron cloud developing when the Ra-Be

or Americium-Beryllium source is
Hydrogen nuclei have a marked
inserted into the soil.
property for scattering and slowing
neutrons. This property is exploited in the Equipments and material :
neutron method for measuring soil water 1. Small fast neutron source such as
content. High energy neutrons (0.1 to 10 radium-beryllium (Mc) or americium-
MeV) emitted from a radioactive beryllium (30 MC)
substance such as radium beryllium 2. Shield for storage of the neutron
(5Mc) or americium-beryllium (30Mc) source. Shielding commonly used
neutron source are called fast neutrons consists of lead, paraffin or
and travel with high speed of the order of polyethylene in a cylindrical shaped
100 miles per second. unit with a cylindrical hole through
When such a fast neutron source is the axis to accommodate the probe.
placed in a moist soils, the emitted 3. Detector of slow neutron - most
neutrons interact with the surrounding commonly used for soil moisture
medium. They collide with the nuclei of measurement is a Bf3-enriched
the soil in a billiard ball fashion, changing proportional counter mounted in a
direction as a result and loosing energy. cylindrical arrangement with
With the energy loses, the speed transistorised preamplifier mounted in
diminishes until it approaches the speed the cable end.
characteristics for molecules at the 4. Counting device (scaler) - the density
prevailing temperature. Such neutrons are of slow neutrons in the soiil and in the
called thermal or slow neutrons. The slow counting tube is evident in the form of
neutrons are finally absorbed by other a given counting rate. This counting
nuclei + and thus their existence ends. In rate may be determined by a rate
a material containing appreciable meter or by a scaler. The first
hydrogen a neutron after the first the indicates the count rate directly and
collision with a hydrogen nucleus is not the second register the total number of
likely to travel much farther consequently counts over a given time period. In
if the neutron source is enclosed in a either case identical results are
material that is rich in hydrogen, it will be obtained.
enveloped in a dense spherical cloud of 5. Access tubing and soil auger sted or
slow neutrons. The density of this cloud aluminum tubing is most commonly
represents an equilibrium between the rate used, but other materials such as
of thermalization and absorbed by the plastic have been used. Two sizes of
medium and the rate of production by the access tubes are in common use. 20-
source. This equilibrium is reached in a gauge steel or aluminum tubings
fraction of micro second after the 1.625 inches (4.13) or 2 inches (5cm)
insertion of source. If the medium outer diameter or 1.9 inches (4.83 cm)
surrounding the source contains less of inner diameter of aluminum
hydrogen, the cloud of slow neutrons will irrigation tubing. Aluminum is
be less dense and extend farther from the practically transparent for fast and
source. This is so because a fast neutron slow neutrons and does not corrode
has to travel further, on an average, to in seriously. Tube once installed may
counter a hydrogen nucleus and start give years of service.
becoming thermalized. Most of the A soil auger slightly smaller the
nitrogen in soil is associated with water tubing should be available for drilling
and lesser amounts with organic matter. the access holes. Moisture probe
Thus, the measurement of soil water by should be calibrated for particular
neutron method is essentially a material and size of access tubing to
measurement of the density of the slow be used.


6. Cable - A 10 meter electric cable Divide readings by standard to

connects detector and scaler. All obtain a count ratio (referred to as relative
possible care should be taken to keep count ratio to standard or per cent of
the connections and firm and the standard etc.) and refer to instrument
connectors free of dirt and moisture. calibration curve to obtain water content
Procedure : by volume at various depths. The
calibration curve supplied with instrument
The access tube is first inserted usually may be used. But, since wide
into the soil after drilling a hole with the differences of soils are know to exist, the
help of auger taking care that no bend in calibration checks, should be made to
the access tube is created. The access tube each soil type as guided by experience in
is kept few inches above the soil and area. It is very important to realize that
covered with an inverted can to prevent the potential precision of the neutron
entrance of trash. method is greater than that of any other
The neutron probe must be method under field condition. Therefore,
inserted in the access tube and held at the calibration should be carried out with
desired depth. Holding the probe may be large homogeneous masses of soil of
done by various means, but a simple and which the bulk density and moisture
effective method is to use an ordinary content (by gravimetric method) are
marking tape, coloured cloth adhesive accurately known.
tape or surgical tape. While making a
References :
measurement turn on the scaler a few
minutes earlier to warmup (transistorized Belchev, D.J. (1952). The measurement
units require no warmup). Make several of soil moisture and density by
standardization counts with the probe neutron and gammaray scattering.
each time. The normal counting time is Highway Res. Board Spl. Rpt. 2 :
minute. The background "thus obtained 98-110.
should not be much more than 100 counts Bouyoucos, G.J. (1937). Evaporation
per minute. As measurements are being water with burning alcohol as a
taken, the standard count is determined rapid means of determining
again from time to time. The frequency moisture content of soils. Soil Sci.
can depend on convenience, plolayout and 44 : 337-383.
experience. It is often convenient to make VanBavel, C.H.M. (1958). Measurement
a standard count at the start and end a of soil moisture content by the
series of readings in each access tube. neutron method. Agr. Res. Serv.
Keep a record of standard count to ARS. 41-24.
provide an index of equipment condition.
After determining the standard count, take
the reading at successive depth intervals
starting at least 18 to 25 cm from the soil
surface. Approximately, 9-15 cm soil
layer is characterized by a single
measurement.


Determination of Soil pH and Electrical Conductivity

S.D. Sawarkar
Determination of pH is actually a water and dilute to 1 litre. Add 1 ml of

measurement of hydrogen ions activity in chloroform or a crystal of thymol per litre
soil – water system. It is defined as as a preservative).
negative logarithm of the hydrogen ion Procedure :
activity. Mathematically, it is expressed (a) Soil to water ratio of 1:2 (pH2)
as: Take 20 g soil in 100 ml beaker and
pH = - log a H+ add 40 ml. of distilled water to it. The
The pH value of a soil is an suspension is stirred at a regular interval
indication of soil reaction i.e. acidic, for 30 minutes. Determine the pH by
neutral or alkaline. The nutrient immersing electrodes in suspension. For
availability is governed by soil reaction. It soils containing high salts, the pH should
is maximum at neutral pH and decreases be determined by using 0.01M calcium
with increase in acidity or alkalinity. chloride solution. (Dissolve 0.110 g of
Thus, pH value gives an idea about the CaCl2 in water and dilute to 1 litre).
availability of nutrients to plants. (b) Saturates soil paste (pHs)
Principle : Add small amount of distilled
water to 250g of air dried soil. Stir the
The pH is usually measured by pH mixture with a spatula. At saturation, the
meter, in which the potential of hydrogen soil paste glistens and flows slightly when
ion indicating electrode (glass electrode) the container is tapped it slides freely and
is measured potentiometrically against ensures cleanly off the spatula. After
calomel saturated reference electrode mixing, allow the sample to stand for an
which also serves as salt bridge. Now a hour. If the paste has stiffened markedly
days, most of the pH meters have single or lost its glistening, add more water or if
combined electrode. Before measuring the free water has collected on the surface of
pH of the soil, the instrument has to be the paste, add an additional weighed
calibrated with standard buffer solution of quantity of dry soil and mix it again. Then
known pH. Since, the pH is also affected insert the electrode carefully in the paste
by the temperature, hence, the pH meter and measure the pH.
should be adjusted to the temperature of (c) Saturation extract (pHe)
the solution by temperature correction The soil is extracted using vacuum
knob. extractor and the pH is measured in the
Reagents : saturation extract.
Standard buffer solutions: These Categories of soil pH values :
may be of pH 4.0, 7.0 or 9.2 and are Soil pH : Interpretaion
prepared by dissolving one standard
< 5.0 : Strongly Acidic
buffer tablet in 100 ml distilled water, It is
necessary to prepare fresh buffer solution 5.1 – 6-5 : Slightly Acidic
after few days. In absence of buffer tablet, 6.6 – 7.5 : Neutral
a 0.05 M potassium hydrogen phthalate
solution can be used which gives a pH of 7.6 – 8.0 : Mild Alkaline
4.0 (Dissolve 10.21 g. of A.R. grade > 8.0 : Strongly Alkaline
potassium hydrogen phthalate in distilled


Electrical Conductivity : Reagents :

Amount of soluble salts in a Potassium chloride: Dissolve
sample is expressed in terms of the 0.7456g dry potassium chloride (AR) in
electrical conductivity (EC) and measured distilled water and make up the volume to
by a conductivity meter. The instrument one litre.
consists of an AC solubridge or electrical Procedure :
resistance bridge and conductivity cell
having electrodes coated with platinum Take 20 g of soil in 100 ml
black. The Instrument is also available as beaker, add 40 ml of distilled water and
an already calibrated assembly shake intermitantly for 30 minutes.
(Solubridge) for representing the Determine the conductivity of the
conductivity of solutions in dSm-1 (deci supernatant liquid with the help of
Siemen per meter) at 250C. conductivity meter. The electrical
conductivity of saturation extract (E.C.e)
Principle is also determined for salinity ratings.
A simple wheatstone bride circuit
is used to measure EC by null method. EC Effect
The bridge consists of two known and (dS m-1)
fixed resistance r1, r2, one variable- <1 - No deleterious effect on crop
standard resistance r4 and the unknown r3. 1-2 - Critical for salt sensitive crops
The variable resistance r4 is adjusted until 2-3 - Critical for salt tolerant crops
a minimum or zero current flows through >3 - Injurious to most crops
the AC galvanometer. At equilibrium.
r1 r3 r1
= or = r3 X r4
r2 r4 r2
Since conductivity is reciprocal of

receptivity, it is measured with the help of
r3 .


Determination of Soil Organic Carbon

S.D. Sawarkar
The majority of mineral surface soils Apparatus and Reagents :

range from 1.2 to 3.5% organic carbon. 1. 500 ml conical flasks.
Since soil organic matter averages about 2. Pipette
58% carbon, it follows that soils generally 3. Burette
range from about 2 to 6 % organic matter 4. Phosphoric acid 85%.
(% O.M. = %C x 1,724. The factor 1.724 5. Sodium fluoride 2%.
= 100/58). There is also a close 6. Sulphuric acid 96 % containing 1.25
relationship between carbon and nitrogen % Ag2SO4.
in soils. Most organic matter average 7. Standard 1N K2Cr2O7 – 49.04 g/liter.
about 5% nitrogen so that the N : C ratio 8. Standard 0.5 N Fe (NH4)2 (SO4)2.
is 1:11.6. Therefore by multiplying the 6H2O 196 g in 800 ml water
soil organic matter percentage by 0.05 an containing 20 cc H2SO4 and diluted
approximate value for the soil nitrogen, to1 litre.
percentage is obtained. 9. Diphenylamine – 0.5g in 20cc water
In soil the chief source of some of and add 100 ml conc. H2SO4.
the nutrients essential for plant growth is
organic matter, such nutrients are N, S Procedure :
and boron is also largely derived form
organic matter. 1. Weigh 1g soil sample in 500 ml
conical flask. Add 10 ml of 1 N
Principle : K2Cr2O7 and 20 ml conc. H2SO4
(containing Ag2SO4). Mix thoroughly
A suitable quantity of the soil is and allow reaction to proceed for 30
digested with chromic acid and Sulphuric minutes.
acid making the use of heat of dilution of 2. Dilute the reaction mixture with 200
Sulphuric acid soil is digested and organic ml water and 10 H3PO4 add 10 ml of
matter of the soil is oxidized. Excess of NaF solution and 2 ml of
chromic acid left over unreduced by the diphenylamine.
organic matter of the soil is determined by 3. Titrate the solution with standard FAS
a titration with standard Ferrous to a brilliant green colour. A blank
Ammonium sulphate solution using without soil should be run
diphenylamine as indicator. simultaneously.
In this exercise, chromic acid in
Observations & Results :
the presence of excess H2SO4 is to be
used as an oxidizing agent for oxidizable Weight of sample - 1g
organic matter of the soil. The heat of Normality of K2Cr2O7 used - 1N
dilution of H2SO4 works as a standardized Vol. of K2Cr2O7 - 10 ml
ferrous sulphate solution. Normality of FAS - 0.5 N
10 0.003 x 100
6+ +++ 4+
OC (%) = Blank (Blank - Reading) X Wt. of soil
4 Cr + 3C 4Cr + 3C
2H2Cr2O7+ 3C+6H2SO4 2Cr2 (SO4)3+ 3CO2+8H2O
K2Cr2O7 + 4 H2SO4 K2SO4+ 2Cr2 (SO4)3+ H2O+3O
Limits :
2FeSO4 + H2SO4+O Fe2 (SO4)3 + H2O Low : < 0.5%
Medium : 0.5 – 0.75%
High : > 0.75%


Determination of Available (Mineralization) Nitrogen in Soil by Alkaline

Permanganate Method
A.K. Upadhyay and Rakesh Sahu
Preparation of soil sample : Principle :
The soil samples from definite depth are A known weight of the soil is
randomly collected from the field with the mixed with alkaline potassium
help of screw auger. All the possible permanganate (KMnO4) solution and
technical precautions as prescribed for distilled. The organic matter present in
standard soil sampling are also taken. soil is oxidized by the nascent oxygen,
Samples are then brought to the liberated by potassium permanganate, in
laboratory, air-dried in the shade and the presence of sodium hydroxide and the
grounded by wooden roller, thereafter released ammonia is condensed and
sieved through 2 mm stainless steel sieve absorbed in known volume of a boric acid
and stored in polythene bags and used for with mix indicator to form ammonium
chemical assay. borate, the excess of which is titrated with
a standard sulphuric acid.
Reactions involved:
I. Distillation :
Alkaline
2 KMnO4 + H2O 2 MnO4 + 2KOH + 3O-
medium (Nascent oxygen)
Oxidation
R.CHNH2COOH + O- R.CO.COOH + NH3
Organic- N fraction (Ammonia)
Distillation
NH3 + H2O NH4OH
Absorption
3NH4OH + H3BO3 (NH4)3BO3 + 3H2O
(Green colour)
II. Titration
2(NH4)3BO3 + 3H2SO4 2(NH4)3SO4 + 2 H3BO3
(Pink colour and original)
Equipment and apparatus :  Automatic Distillation System
(Model Classic DX): It is fully
1. KEL PLUS Automatic Nitrogen
automatic distillation system with
Estimation System
programmable auto run digital
The said instrument is used for
features, with automatic dilution and
determination of available nitrogen in
addition of boric acid, NaOH and
soil. It consists of the following:
KMnO4. Both modes (auto and


manual) are available for distillation  With the absorption of ammonia, the
reagents addition. pinkish colour turns to green.
 Refrigerated Water Cooling  Nearly 150 ml of distillate is
System for Condenser (Model Kel collected in about 10 minutes.
Freeze): It is refrigerated water  The green colour distillate is titrating
cooling system for distillation and with 0.02N sulphuric acid and the
condensing system with inbuilt colour changes to original shade
compressor and recirculator pump. (pinkish color).
2. Electronic balance  Simultaneously, blank sample
3. Burette (without soil) is to be run.
4. Conical flask  Note the blank & sample titer reading
5. Distilled water (ml) and calculate the available
Reagents : nitrogen in soil.
1. 0.32 % potassium permanganate
(KMnO4) solution. Calculations :
R (Titer reading - Blank reading)
2. 2.5 % sodium hydroxide (NaOH). × Normality of acid
3. 2 % boric acid solution containing 20 Available N × Atomic weight of nitrogen ×
- 25 ml of mixed indicator / liter. (kg ha-1) = Weight of one hectare of soil
Sample weight (g) x 1000
4. Mixed indicator: 0.066g methyl red +
R × 0.02 × 14 × 2.24 × 106
0.099g bromocresol green dissolve in =
5 x 1000
100 ml of 95 % alcohol.
5. 0.02 N sulphuric acid (H2SO4). Factor = R x 125.44
Procedure : Interpretation of results :

 Weigh 5 g of prepared soil sample Available N (kg ha-1) Soil rating
and transfer it to the digestion tube. < 280 : Low
 Load the tube in distillation unit and 280-560 : Medium
other sides of hose keep 20 ml of 2 %
boric acid with mixed indicator in > 560 : High
250 ml conical flask. References :
 25 ml each of potassium
permanganate (0.32 %) and sodium Subbiah, B.V. and Asija, G. L. (1956). A
hydroxide (2.5 %) solution is rapid procedure for the estimation
automatically added by distillation of nitrogen in soils. Curr. Sci., 25:
unit programme. 259-260.
 The sample is heated by passing
steam at a steady rate and the
liberated ammonia absorbed in 20 ml
of 2 % boric acid containing mixed
indicator solution kept in a 250 ml
conical flask.

17

Determination of Total Nitrogen in Soil and Plant

A.K. Upadhyay and Rakesh Sahu
Total nitrogen is estimated by the micro- The micro-Kjeldahl method consists of

Kjeldahl method as per procedure the three steps;
suggested by AOAC (1995). 1. Digestion
2. Distillation and
Preparation of plant and soil samples : 3. Titration.
The plant analysis has been Equipment and apparatus :
considered as a superior diagnostic 1. KEL PLUS Automatic Nitrogen
technique for mineral content. Whole Estimation System :
plant is dried in open air for few days
after that it was further dried in hot air The said instrument is used for
oven at about 60 ± 2o C for eight to ten determination of nitrogen. It consists of
hours per day to achieve complete drying. the following;
After drying, whole plant is powdered  Macro Block Digestion System
with the help of a grinder, passed through (Model KES 12L): This digestion system
2 mm stainless steel sieve and used for is suitable for soil, plant, water,
chemical assay. The soil sample from pesticides, fertilizers, food and feed
definite depth was randomly collected samples. It is microprocessor based
from the field with the help of screw automatic twelve place macro block
auger. All the possible technical digestion system with temperature
precautions as prescribed for standard soil controller fitted with sensor break
sampling were also taken. Samples were protection (Microprocessor based) feature
brought to the laboratory, air-dried in the and temperature range from 50-450 0C.
shade and grounded by wooden roller,  Acid Neutralizer Scrubber (Model
thereafter sieved through 2 mm stainless KEL VAC): It is used to neutralize the
steel sieve and stored in polythene bags acid fumes, which are absorbed in 15%
and used for chemical assay. sodium hydroxide and dissolved in water
stored in the system tank. After every 2
Principle : cycles of digestion, the 15% sodium
Nitrogen in samples like plant and hydroxide solution is replaced and after 3
soil exists in a very complicated bonding cycles of digestion, acid fumes dissolved
structure. During digestion, a known in water tank is drained off and refilled
weight of the plant/soil samples in the with fresh water in the system tank.
presence of sulphuric acid with catalyst  Automatic Distillation System
mixture under high temperature is (Model Classic DX): It is fully automatic
digested where complicated structures are distillation system with programmable
broken to simple structure, thereby auto run digital features, with automatic
releasing nitrogen in the form of dilution and addition of boric acid and
ammonium radical (NH4+). During NaOH. Both modes (auto and manual) are
distillation in presence of sodium available for distillation reagents addition.
hydroxide, the released ammonia is  Refrigerated Water Cooling
condensed and absorbed in known System for Condenser (Model Kel
volume of a boric acid with mix indicator Freeze): It is refrigerated water cooling
to form ammonium borate, the excess of system for distillation and condensing
which is titrated with a standard sulphuric system with inbuilt compressor and
acid. recirculator pump.


2. Electronic balance liberated ammonia absorbed in 20 ml

3. Burette of 4 % boric acid containing mixed
4. Pipette indicator solution kept in a 250 ml
5. Conical flask conical flask.
6. Measuring cylinder  With the absorption of ammonia, the
7. Distilled water pinkish colour turns to green.
Reagents :  Nearly 150 ml of distillate is
1. Concentrated sulphuric acid (H2SO4). collected in about 8 minutes.
2. Catalyst mixture: Mix with 250 g  Simultaneously, blank sample
potassium sulphate (K2SO4), 50 g (without plant/soil) is to be run.
cupric sulphate (CuSO4. 5 H2O) and III. Titration :
5 g metallic selenium powder in the
ratio of 50:10:1.  The green colour distillate is titrating
3. 40 % sodium hydroxide (NaOH). with 0.02N sulphuric acid and the
4. 4 % boric acid containing 20 - 25 ml colour changes to original shade
mixed indicator /liter. (pinkish color).
5. Mixed indicator: 0.066 g methyl red  Note the blank & sample titer reading
+ 0.099 g bromocresol green dissolve (ml) and calculate the total nitrogen
in 100 ml of 95 % alcohol. content present in plant/soil samples.
6. 0.02N sulphuric acid (H2SO4). Calculations :
Procedure : R (sample titer-blank titer) x
Normality of acid x Atomic
I. Digestion : Nitrogen content weight of nitrogen x 100
in plant (%) =
 Weigh 0.5 g of prepared plant sample Sample weight (g) x 1000
or 1 g of soil sample and transfer it to R x 0.1 x 14 x 100
=
the digestion tube. 0.5 x 1000
 Add 10 ml of concentrated sulphuric Factor = R x 0.28
acid and 5 g of catalyst mixture to the R (sample titer-blank titer) x
sample. Normality of acid x Atomic
Nitrogen content weight of nitrogen x 100
 Load the digestion tubes in to the in soil (%) =
Sample weight (g) x 1000
digester and heat the digestion block.
 Switch on the digestion unit and set
R x 0.1 x 14 x 100
the initial temperature 100 0C till =
1 x 1000
frothing is over.
 Then block temperature is raised to Factor = R x 0.14
400 0C. The effective digestion starts Crude protein content :
only at 360 0C and beyond 410 0C.
The total nitrogen is estimated by
 The sample turns light green colour micro-Kjeldahl method as per procedure
or colorless at the end of the suggested by AOAC (1995) and the crude
digestion process. protein is calculated by the following
II. Distillation : formula:
 After cooling the digestion tube, load Crude protein content (%) = micro-Kjeldahl
the tube in distillation unit and other nitrogen content (%) x 6.25 (based on the
side of hose keep 20 ml of 4 % boric assumptions that nitrogen constitutes 16 % of
protein).
acid with mixed indicator in 250 ml
conical flask. References :
 40 ml NaOH (40 %) is automatically AOAC, (1995). Official Methods of Analysis.
added by distillation unit programme. 16th edn. Association of Official
 The digested sample is heated by Analytical Chemists, Washington,
passing steam at a steady rate and the DC.


Determination of Phosphorous in Soil and Plant

R.K. Thakur
The phosphorus is an essential plant  Above both solution mix

nutrient and it occurs in many different thoroughly and made one litre in
forms. Therefore, a reliable procedure for volumetric flask with the help of
measuring the amount both in soil as well distilled water.
as in plant is needed. There are many  Add these dissolved reagents to
methods available for the determination, one litre of 5N H2SO4.
however, colorimetric measurement is
5. Ascorbic acid working solution
presented here:
(Reagent B): Dissolve 1.056 g of
Principle : ascorbic acid in 200 ml of reagent A
and mix. This ascorbic acid (reagent
Phosphorus is extracted from the B) should be prepared as required
soil with 0.5 m NaHCO3 at a nearly because it does not keep more than
constant pH of 8.5. The phosphate ion in 24 hours.
solution treated with ascorbic acid in an
acidic medium provides a blue colour 6. Standard phosphate solution:
complex. Measurement of the quantitative Weigh 0.4393 g of potassium
determination of phosphorous in soil dihydrogen phosphate (KH2PO4) into
(Olsen’s et al., 1954) one litre volumetric flask. Add 500
ml of distilled water and shake the
Reagents : contents until the salt dissolves.
Dilute the solution to one litre with
1. 0.5 M Sodium bicarbonate distilled water to get 100 ppm P
(NaHCO3) solution: Dissolve 42 g solution. Dilute 20 ml of 100 ppm P
of NaHCO3 in distilled water to get solution to one litre to get form-
one litre solution and adjust the pH of working solution of 2 ppm.
the solution to 8.5 by small quantity
of NaOH. Preparation of standard curve :
2. Activated Charcoal: Darco G-60 (P-  Take different concentration of P (0,

Free) 1, 2, 3, 4, 5, etc ml of 2 ppm standard
3. 5 N Sulphuric acid (H2SO4) P Solution) in 25 ml volumetric
Solution: Add 141 ml of con. H2SO4 flasks.
to 800 ml of distilled water. Cool the  Add 5 ml of the 0.5M NaHCO3
solution and dilute to one litre with extracting solution to each flask, and
distilled water. acidify with 5N H2SO4 drop by drop.
4. Reagent A:  Add about 10 ml distilled water and 4
 Dissolve 12.00 g of ammonium ml of reagent ‘B’, then shake the
paramolybdate in 250 ml of solution.
distilled water.  Make the volume 25 ml by distilled
 Dissolve 0.2908 g of potassium water.
antimony tartrate (KSbO.C4H4O6)  The intensity of blue colour is read
in 100 ml distilled water. on spectrophotometer at 660 nm
wavelengths after 10 minutes.


 Plot the curve by taking P Determination of total phosphorus in

concentration on X axis and plant :
colorimeter reading on Y axis. Repeat
Principle : Vanadate molybdate and
the process till you get straight line
orthophosphates react to give a yellow
relationship.
colour complex in acidic medium. The
 Calculate the factor i.e. 1 colorimeter intensity of colour provide the basis of
reading is equal to how much ppm of quantitative measurement of total P in
phosphorus? plant (Koenig and Johnson, 1942).
Procedure : Apparatus and reagents :
 Take 2.5 g of soil sample in 150 ml  Colourimeter/spectrophotometer
conical flask and 0.5 g Darco G-60
activated charcoal.  50 ml volumetric flask
 Then add 50 ml of 0.5 M NaHCO3  ammonium molybdate ammonium

solution and shake the solution for 30 vanadate (in NHO3) solution :
minute in a shaker. Similar processes Dissolve 2.5 g (NH4)6 Mo7O2.4 4H2O
run for a blank without soil. in 400 ml distilled water. Dissolve
1.25 g of ammonium vanadate in 300
 Filter the suspension through the ml boiling water. Add the ammonium
Whatman no. 40 paper. vanadate solution to the ammonium
 Take 5 ml aliquot of the extract in a molybdate solution and cool to room
25 ml volumetric flask, and acidify temperature. Add 250 ml conc. NHO3
with 5N H2SO4. and dilute to 1 lit.
 Add small quantity of distilled water,  Phosphate standard solution : Dissolve
and then add 4 ml of reagent B. 0.2195 g KH2PO4 and dilute to 12%.
This solution contains 50 µg
 The intensity of blue colour is read phosphorus/ml.
on spectrophotometer at 660 nm
wavelengths after 10 minutes. Procedure :
Observations : Preparation of standard curve :
1. Weight of soil sample : 2.5 g  Transfer 0, 1, 2, 3, 4 and 5 ml of 50
2. Volume of extractant used : 50 ml ppm P solution to 50 ml volumetric
flasks in order to get 0, 50, 100, 150,
3. Volume of filtrate used : 5 ml
200 and 250 µg P.
4. Absorbency : R
 Add 10 ml vanadomolybdate reagent
5. Absorbency from standard curve : A
make up the volume and mix the
6. Concentration of P for absorbency A : B ppm content thoroughly.
Calculation :  Read the transmittance/absorbance at
R x F x 50 x 2.24 420 mµ (blue filter).
Available P (kg ha-1) =
5 x 2.5  Plot the reading against µg P and
Where, F (factor) = B / A calculate the factor (F).
Digestion of plant material :
Limits of available P in soil :
Very low : Less than 5 P kg ha-1 Take one gram of plant material in
Low : 5-10 P kg ha-1 digestion flask. Add 10-15 ml of Diacid
Medium : 10-20 P kg ha-1 (3:1: Nitric acid : Perchloric acid) mixture
High : 20-40 P kg ha-1 and swirl the content in 150 ml
volumetric flask. Place the content on hot
Very high : More than 40 P kg ha-1


plate till the digestion is over. Filter the Reference :

solution in 100 ml conical flask, wash the
Koenig, R.A. and Johnson, C.R. (1942).
residue on filter paper several times with
Colorimetric determination of
the hot water. Make up the volume with
biological materials Ind. Eng.
distilled water, store the solution in air
Chem. Analyt. Edn. 14 : 155-156.
tight container.
Olsen, S.R., Cole, C.V., Watanabe, F.S.
Estimation : and Dean, L.A. (1954). Estimation
 Transfer 10 ml dilute in 50 ml of available phosphorus in soils by
volumetric flask. extraction with sodium
bicarbonate. Circ. U.S. Dept.
 Add 10 ml ammonium molybdate
Agric. 939 : 1-19.
vanadate solution shake the content.
 Make up the volume and record the
reading as per the procedure under
preparation of standard curve.
Calculation :
50 µg = R
1R = 50/R µg (Factor)
Factor (F) x Reading x 100x100

Total (%) sample
P = ------------------------------------------
10000 x 1000 x 10 x 1


Determination of Potassium in Soil and Plant

S.S. Baghel
The available potassium i.e. H2O and volume to be made one litre and
exchangeable and water soluble then adjust the pH 7.0 .
potassium is determined by extracting soil (b) Standard Potassium Solution :
with neutral normal ammonium acetate Dissolve 1.9066 g of dried KCl
solution. The estimation of potassium is (AR) in distilled water and dilute to one
carried out by flame photometer. litre. This is 1000 mg kg-1 K solution.
1. Principle : 100 ml of this solution diluted to 1 lit. to
make 100 ppm K solution.
The principle underlying this is
that a large number of elements when 4. Preparation of the standard curve :
excited in a flame, emit radiation of Take 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and
characteristic wave length. The excitation 10 ml of 100 mg kg-1 K solution in
cause one of the outer electron of neutral different 25 ml volumetric flasks. Make
atoms to move to an outer orbit of higher up the volume with 1N NH4OAc Soln.
energy level or the atoms may be excited Adjust the flame photometer reading at
sufficiently to loose an electron zero with blank (zero K) solution and at
completely from the attractive force of the 100 for 40 mg kg-1 K solution. Take the
nucleus where excited atom return to flame photometer readings for every
lower energy level, light at characteristic dilution. Plot the standard curve on graph
wave length is emitted. Excited atoms or paper by taking K concentration on X axis
ions give line radiation at very definite and flame photometer reading an y axis.
wave length and thus K gives at 404.4 and This will give a factor (F) of 1 flame
767 mµ. The flame photometer photometer reading = 0.4 mg kg-1 K.
employees a relatively low temperature
5. Procedure :
excitation and measures with a photocell
the emission intensity which is Take 5g soil in 100 ml conical
proportional and to concentration in flask and add 25 ml of 1N NH4OAc Soln.
selected wave length (767 mµ) and for Shake the content for 5 minutes and then
this red filter is used. filter through Whatman No.1 filter paper.
2. Apparatus and reagents : Potassium extract is measured by flame
photometer after calibration.
a) Flame photometer with red filter,
6. Calculation :
b) Pipette, volumetric flasks and conical
flask (100 ml). RxFx25x100x20x1.121
Available K (kg ha-1) =
3. Reagents : 5 x 1000
(a) Neutral Normal Ammonium Acetate :
= R x F x 11.217.
Add 58 ml of glacial acetic acid to Limits of available K in soil :
about 600 ml H2O and then add 70 ml of
Very low : Less than 200 K kg ha-1
concentrated ammonia (sp. gr 0.90) Dilute
the solution to one litre. Then adjust pH Low : 200 – 250 K kg ha-1
of solution at 7.0 with the help of Medium : 250 – 400 K kg ha-1
ammonia or acetic Acid or this can be High : 400 – 600 K kg ha-1
prepared by dissolving ammo. Acetate Very high : More than 600 k kg ha-1
(CH3COONH4) (77.08 eq.wt.) directly in


8. Precaution : (b) Determination of K :

a) These should not be any turbidity or Take the aliquot and get the
suspended particles in extract, it will reading of K through flame photometer
chock the capillary feeding tube . using red filter and calculate the amount
b) The gas and air pressure should be of K in the plant sample on the oven dry
constant. matter basis.
c) If sample reading goes beyond 100
then dilute the extract. K (%) in plant sample = X x 4 x 10-3
9. Determination of k in plant sample : References :
(a) Wet digestion : Black, C.A. (1965) Methods of soil
Place 1-2g of ground plant sample analysis Part I Am. Soc. Agron.
in 100ml digestion flask. Add 20-25 ml of Inc. Publi. Madison Wisconsin
acid mixture Acid mixture 750 ml conc. USA.
HNO3 + 150 ml conc H2SO4 + 300 ml of
HClO4 and mix the contents of the flask
by swirling well. Heat the flask at a low
temp and then slowly increase the flame
or temp. of hot plate. Completion of
digestion is confirmed when liquid is
colorless. After cooling, add 20-25 ml
H2O and filter through whatman No.40
into a 100 ml/250 ml volume flask and
make up the volume.


Determination of Sulphur in Soil and Plant

B.S. Dwivedi
Principle : and dilute to one litre with dilute

acetic acid.
Besides some amount in the soil
3. Barium chloride: Pass AR grade
solution, available sulphur in mineral soils
BaCl2 salt through 1 mm sieve and
occurs mainly as adsorbed SO4= ions.
store for use.
Phosphate ions (as monocalcium
4. Standard stock solution (2000 mg S
phosphate) are generally preferred for
L-1): Dissolve 1.089 g of oven dried
replacement of the adsorbed SO4= ions.
AR grade potassium sulphate per 100
The extraction is also carried out using
mL.
CaCl2 solution. However, the former is
5. Working standard solution (10 mg S
considered to be better for more efficient
L-1): Measure exactly 2.5 mL of the
replacement of SO4= ions. Use of Ca salts
stock solution and dilute to 500 mL.
have a distinct advantage over and leads
6. Barium sulphate seed suspension:
to easy filtration SO4= in the extract can
Dissolve18 g of AR grade BaCl2 in 44
be estimated turbid metrically using a
mL of hot water and add 0.5 mL of the
colorimeter/spectrophotometer.
standard stock solution (given above).
A major problem arises when the
Heat the contents to boiling and then
amount of extracted sulphur is too low to
cool quickly. Add 4 mL of gum acacia-
be measured precisely.; To overcome this
acetic acid solution to it. Prepare a
problem, addition of seed solution of
fresh seed suspension for each
known S concentration is added to the
estimation everyday.
extract to raise concentration to easily
7. Dilute nitric acid (approx 25%):
detectable level. Sulphur in the extract
Dilute 250 mL of AR grade conc.
can also be estimated by a colorimetric
HNO3 to one litre.
method using barium chromate (Nemeth
8. Acetic-phosphoric acid: Mix 900 mL
1964; Palaskar et al. 1981), but the
of AR grade glacial acetic acid with
turbidimetric method (Chesnin and Yien
300 mL of H3PO4 (AR grade).
1950) given below is mainly used in the
soil testing laboratories. Procedure :
Instruments : 1. Weight 20 g of soil sample in a 250
mL conical flask.
(i) Colorimeter or spectrophotometer or
2. Add 100 mL of the monocalcium
autoanalyzer.
phosphate extracting solution (500 mg
(ii) Mechanical shaker
P L-1) and shake for one hour. Filter
Reagents through Whatman No. 42 filter Paper.
3. Measure 10 mL of the clear filtrate into
1. Mono-calcium phosphate extracting a 25 mL volumetric flask.
solution (500 mg P L-1): Dissolve 2.035 4. Add 2.5 mL of 25% HNO3 and 2 mL
g of Ca(H2PO4)2.H2O per litre. of acetic-phosphoric acid. Dilute to
2. Gum acacia-acetic acid solution: about 22 mL, stopper the flask and
Dissolve 5 g of chemically pure gum shake well.
acacia powder in 500 mL of hot water 5. Shake the BaSO4 seed suspension and
and filtered in hot condition through then add 0.5 mL of it and 0.2 g of
Whatman No. 42 filter paper. Cool BaCl2 crystals. Stopper the flask and
invert three times and keep.


6. After 10 minutes, invert 10 times and distilled water, store the solution in air
keep. After another 5 minutes, invert 5 tight container.
times. Estimation :
7. Allow to stand for 15 minutes and
Take 10 ml aligust from extract
then add 1 mL of gum acacia-acetic
and proceed as per the method described
acid solution.
under preparation of standard curve
8. Make up the volume, invert three
(Bardsley and Lancaster, 1960).
times and keep aside for 90 minutes.
9. Invert 10 times and measure the
Calculation :
colour intensity at 440 nm (blue
filter).
5 µg = R
10. Run a blank side by side.
1R = R/5 µg (Factor)
Preparation of standard curve for S :
1. Place 2.5,5.0,7.5,10.0,12.5 and 15.0 Total S (%) =
Factor x Sample R x1000 x 100 x 100
mL portions of the working standard -----------------------------------------------
solution (10 mg S L-1) into a series of 1000 x 10 x 1
25 mL volumetric flasks to obtain
25,50,75,100,125 and 150 µg S. References :
2. Proceed to develop turbidity as
Arora, C.L. and Bajwa, M.S. (1994).
described above for sample aliquots.
Curr. Sci. 66 : 314-316.
3. Read the colour intensity and prepare
Bardsley, C.S. and Lancaster, J.P. (1960).
the curve by plotting readings against
Proc. Soil Sci. Soc. Am. 24 : 265.
sulphur concentration (In µg in the
Chesnin, L. and Yien, C.N. (1951). Proc.
final volume of 25 mL)
Soil Sci. Soc. Am. 15 : 149.
Calculation :
R x 100
Available S in soil (mg kg-1) =
10 x 20
Where, r stands for the quantity of S in
mg as obtained on X-axis against a
reading.
Determination of total sulphur in plant:
Sulphur is an essential plant
nutrient and occurs in many different
forms. The procedure for total sulphur
estimation is as follows :
Digestion of plant material :
Take one gram of plant material in
digestion flask. Add 10-15 ml of Diacid
(3:1: Nitric acid : Perchloric acid) mixture
and swirl the content in 150 ml
volumetric flask. Place the content on hot
plate till the digestion is over. Filter the
solution in 100 ml conical flask, wash the
residue on filter paper several times with
the hot water. Make up the volume with


Determination of Zn, Cu, Fe and Mn in Soils and Plants by Atomic

Absorption Spectrophotometer
G.D. Sharma
All atoms can absorb light at certain Beer’s Law : Light absorption is
discrete wavelengths corresponding to the proportional to the number of absorbing
energy requirement of the particular atom. atoms in the sample.
When at ground state the atom absorbs The combined Beer - Lambert law may
light it is transformed into the excited be given as :
state. It is the same atom containing more It = Io – (abc)
energy. This energy is measured in Io
relation to the ground state and a thus log10 --- = abc = absorbance
particular excited state say for example in It
case of Na may be 2.2 eV (electron volts) Where, Io = incident radiation power
above the ground state. It = transmitted radiation power
Each transition between different a = absorption coefficient
electronic energy states is characterized b = length of absorption path
by a different energy and by a different c = concentration of absorbing
wavelength. These wavelengths are atoms
sharply defined and when a range of i.e. the absorbance is proportional
wavelengths is surveyed, each wavelength to the concentration of the elements for a
shows as a sharp energy maximum (a given absorption path length at any given
spectronic line). These characteristic lines wave length.
distinguish atomic spectra. The lines, In principle, it might be possible
which originate in the ground state of to calculate the concentration directly
atom, are most often of interest in atomic from the above equation. In practice,
absorption spectroscopy. These are called however, the a and b are constants hence
the resonance lines. The atomic spectrum, the variation of results is directly related
characteristic of each element, then the concentration of atoms. For analysis,
comprises a number of discrete lines, the absorbance of different concentration
some of which are resonance lines. Most of standard solution is first measured with
of the other lines arise from excited states the help of atomic absorption
rather than the ground state. The lines of spectrophotometer and then the results of
excited states are not useful generally in unknown samples are compared with the
atomic absorption analysis as most of the standards and thus concentration of
atoms in a practical atomizer are found in unknown sample is calculated.
the ground state.
Atomic absorption spectrophotometer :
The relationship of light absorbed
by the atom in ground state and their Atomic absorption spectro-photo-
concentration in the solution is defined in meter is based on the principle that when
the fundamental laws of light absorptions. atomic vapours of an element are
Lambert’s Law : The portion of light irradiated by the radiation of a
absorption by a transparent medium is characteristic wavelength (i.e. the light
independent of the intensity of the from a source whose emission lines are
incidence light and each successive unit those of the element in question), they
thickness of the medium absorbs an equal absorb in direct proportion to the
fraction of the light passing through it. concentration of the element being
determined.

Instrument features : switching and calculation of average and

standard deviations in repetitive runs.
A wide range of atomic absorption
spectrophotometer is available today, all Collection and preparation of soil and
of them have the basic features in plant samples : To avoid contamination,
common and consist of the following soil samples are to be collected in plastic
components: tub, using rust free instrument or wood
(a) A Light source : and kept in polythene lined cloth bags.
Samples are prepared with the help of
A Light source emits the spectrum wooden mortar and pastle and sieved
of the element to be determined. The most through 2mm nylon screen/mosquito net
widely used light source is hollow cloth or stainless steel sieve.
cathode lamp which is designed and Similarly plant samples (leaves,
operated in such a way that the lines to be grains or straw) should be washed with
measured are sharp, of stable intensity 0.01N HCl, rinsed with glass distilled
and free from background. water dried in oven at 65°C and crushed
(b) Atomizer-Burner assembly : with the help of stainless steel scissors.
A means of producing atomic Soil extraction : DTPA offers the most
vapours of the element to be analyzed. favourable combination of stability
The solution to be analyzed is drawn by constants for the simultaneous
capillary and converted into stream of complexing of Zn, Cu, Fe and Mn, Cd,
compressed air to a fine spray which after Co, Ni and Pb (Lindsay and Norvell,
condensation of larger droplets is mixed 1978). Buffering of extractant in a slightly
with the fuel gas acetylene and burnt in a alkaline pH range (7.3) by including
long flame (at 2100-2400oC) in a stainless soluble Ca2+, avoids the dissolution of
steel burner. CaCO3 with the release of occluded
(c) A Monochromator : micronutrients due to CO2 partial pressure
It isolates the absorbing of approximately 10 times that in
resonance lines from other non absorbing atmosphere, as the soil contains slightly
lines. When the light coming from the higher CO2 levels than found in the
HCL, after traversing the flam, enters the atmosphere.
monochromator which is already set at the (a) Extracting solution : (0.005 M
wavelength of the resonance lines of the DTPA) Dissolve 1.9679g of DTPA
desired element, the monochromator (Diethylene tri amine penta acetic
performs its function. acid) + 13.3 ml TEA (Triethanol
(d) A Detector : amine) + 1.47g CaCl2 .2H2O in 200
ml distilled water, dilute to 900 ml,
It measures the magnitude of adjust pH 7.3 with 6N HCl while
absorption of the characteristic radiation. stirring and then make upto 1 liter
(e) A Photomultiplier Tube : and mix thoroughly.
It amplifies the absorption signal (b) Apparatus required : Shaker
and converts the light radiation into (Horizontal or Rotatory), iodine value
electrical energy. flasks (100 ml capacity) or conical
(f) A readout system : flasks with glass stoppers, funnels,
It measures the absorbance in filter paper whatman No.1, plastic
volts. It is normally a strip chart recorder, storage bottles and Atomic absorption
a digital display, a meter or printer. The spectrophotometer.
presently available AAS have features (c) Stock Standard Solutions : The
like automatic calibration with one or standard solutions of different micro-
more standards, automatic curve nutrients should preferably be
corrections, automatic and foolproof gas

prepared by using their wires. pass through the small aperture, forming a
Dissolve 1g wire in a minimum defined area of high excitation and hence
volume of 1:1 nitric acid and dilute to high light emission. A suitable window
1000ml with distilled water to obtain transmits the light to the spectrometer's
1000 µg/ml solution of micro-nutrient, optical system.
or take salts of metals as follows: To obtain successful background
Zn- 4.398g l-1 ZnSO4,7H2O correction the deuterium lamp must be
Cu- 3.929g l-1 CuSO4,5H2O correctly aligned, and its intensity must be
Fe- 4.977g l-1 FeSO4,7H2O matched to that of the hollow cathode
Mn- 3.598g l-1 MnSO4,H2O. lamp.
The prepared standards are also It is important that both the
available in the market. Out of these deuterium source and the hollow cathode
standards, prepare working solution of source are aligned to follow the same
50 ppm. Then a series of standard optical path. If they are not, then the two
solution of 0.5, 1.0, 1.5, 2.0 and 2.5 measurements may not be made on the
ppm may be prepared for each metal. same atom population and significant
errors may occur.
(d) Background correction : The In order to balance the intensity of
reading of a spectral line always the deuterium lamp with the hollow
includes any contribution from the cathode lamp, it may be necessary to
flame and sample matrix. Failure to change the hollow cathode lamp current
correct properly for the background to a higher or lower value depending on
reading can be a source of serious the relative intensities of the lamps.
error. Although the need for fast Although most modern AA spectro-
background correction is most photometers incorporate so called
obvious with graphite furnace work, "simultaneous" background correction, they
it is also a consideration with flame rely on two measurements separated slight
atomic absorption. in time. One measurement is of the total
The most common method of absorbance (atomic plus background) and
background correction in atomic the other is of the background only. The
absorption spectrometry involves the use background is electronically subtracted
of a continuum source such as a from the absorbance to give the background
deuterium lamp to measure the corrected atomic absorbance with the
background. The source used is a continuum source method of background
deuterium filled discharge lamp, which correction, the total absorbance is measured
emits an intense continuum spectrum during the hollow cathode lamp pulse and
from 190 nm to about 400 nm. This is the the background during the deuterium lamp
region where most atomic absorption pulse. With the Zeeman method using a
lines occur and where the effects of modulated magnetic field, the total
background absorption are most absorbance is measured with the magnetic
field off and the background with the field on.
pronounced. The poly-atomic gas D2, is
used in the lamp because a continuum is (e) Soil analysis : Weigh 12.5g soil
produced rather than a line spectrum. sample in 100 ml iodine value flasks.
The deuterium lamp is different from Add 25 ml DTPA solution. Shake
a hollow cathode lamp in construction and this mixture for 2 hours on shaker at
operation. The lamp incorporates a 70 to 80 oscillation per minute, filter
heated, electron-emitting cathode, a metal through acid washed distilled water
anode and a restrictive aperture between rinsed, whatman No.1 filter paper and
the two. A discharge current of several collect the filtrate in plastic bottles.
hundred milli amperes excites the Determine the content of
deuterium gas. The discharge is forced to micronutrients on atomic absorption
spectrophotometer.


(i) Plant analysis : Weigh 0.5g plant Factors : For soil multiply the
sample in a conical flask (corning, concentration read on AAS computer
100 ml capacity). Add 10 to 12 ml of sheet by “2”. Similarly for plants the
di acid mixture (1 perchloric + 4 multiplying factor will be 100 to get
nitric acid) and digest the mixture on concentration in mg kg-1.
hot plate till the residue is colour less.
Now take off, cool dilute with
Reference :
distilled water and filter through
whatman No.1 filter paper. Make up Lindsay, W.L. and Norvell, W.A. (1978).
the volume of digestate to 50 ml. Proc. Soil Sci. Soc. Am. 42 :
Read for micronutrient content on 421-428.
atomic absorption spectro-
photometer.


Estimation of Boron in Soils, Plants and Water

G.D. Sharma
Boron occurs as anion in soils and is  In alkaline soils the availability of B is

required by plants in very small quantity. high and may be even toxic for plant
Water soluble B makes the estimate of its growth.
availability to plants. Total boron in soils Besides this the low moisture
varies from 20 to 200 mg kg-1 and availability also causes B deficiency.
available (water soluble) boron in soils Irrigation water containing Boron
ranges from 0.03 to 12 mg kg-1 between 0.3 to 0.6 mg kg-1 can be used
respectively. The threshhold value safely, whereas, irrigating soils with water
ranging from 0.1 to 0.5 mg kg-1 (water containing 1 to 3 mg kg-1 B causes
soluble B) depends upon the soil type, toxicity of B in plants.
crops, and other factors, below which the Boron determination (Azomethine H
response to applied boron may be Method) :
expected. Some sensitive crops to boron
deficiency are listed in table 1. Its Azomethine H forms coloured
availability is affected by soil pH as complex with H3BO3 in aqueous media.
under: Over a concentration range of 0.5 to 10
 Deficiency of B is generally observed µg B/ml the complex is stable at pH 5.1.
in old acid leached soils. Maximum absorbance occur at 420 nm
 Availability increased with the rise in with little or no interference from a wide
soil pH having significant positive variety of salts. This technique is rapid,
correlation with pH rising from 4.7 to reliable and more convenient to use than
6.7. traditional procedures employing carmin,
 In neutral, saline and calcareous soils curcumin or quinalizarin (John et al.,
the B availability again decreases with 1975).
the rise in soil pH having significant Apparatus :
negative correlation with the rise in
pH from 7.1 to 8.1. In calcareous soils (1) Spectrophotometer
B fixation occurs with the (2) Poly-propylene tubes 10 ml capacity.
condensation of borate radical into Reagents :
long chains in the presence of Ca. 1. Distilled water
Table 1 : Sensitivity of crop to Boron 2. Buffer solution : Dissolve 250 g of
deficiency ammonium acetate (NH4OAc) and 15
Sensitive Medium Low g of ethylenediaminetetracetic acid
Alfalfa Apple Barley (EDTA disodium salt) in 400 ml of
Cauliflower Cabbage Beans distilled water. Slowly add 125 ml of
Rape seed Carrot Corn glacial acetic acid and mix.
Conifers Clover Grasses 3. Azomethine H reagent : Dissolve 0.45
Peanuts Cotton Oat g of azomethine H in 100 ml of 1% L
Sugarbeet Onion ascorbic acid solution. Fresh reagent
Turnip Pea should be prepared weekly and stored
Potato in a refrigerator.
Soybean 4. Calcium hydroxide suspension : Add
Wheat 0.4g Ca(OH)2 to 100 ml distilled
Rice water.


5. 0.1 N HCl : Add 8.3 ml conc. HCl to Transfer 20 ml aliquot to

900 ml distilled water, mix, cool to evaporating dish, add 2 ml Ca(OH)2)
room temperature and make up the suspension and evaporate the solution to
volume to 1000 ml. dryness. Heat the evaporating dishes
6. Calcium chloride 0.01 M Dissolve gently to destroy organic matter, cool to
1.11 g of anhydrous CaCl2 in 900 ml room temperature, add 5 ml 0.1N HCl.
distilled water and make up the Triturate the residue with rubber
volume to 1000 ml. policeman to ensure the complete
7. Boron standard solution : Dissolve dissolution of the residue (Bingham,
0.114g of Boric acid (H3BO3) in 1982).
distilled water and adjust the volume For analysis of B pipette 1 ml of
to 1000 ml. Each ml contains 20 µg B. the aliquot and proceed as for the standard
Dilute 10, 20, 30, 40 and 50ml of the curve.
stock solution to 100 ml with distilled 2. Plant digest : Take 0.5 g plant
water to have solution with B sample in porcelain/platinum dishes Add
concentration of 2,4,6,8 and 10 µg of 0.5 g Ca(OH)2. Ignite the sample in the
B/ml respectively. Include a distilled muffle furnace at 550°C for 4 hours to
water sample for the 0.0 µg of B/ml obtain white grey ash. Cool the dishes and
standard solution. moist the ash carefully with little distilled
Procedure : water and then add 5 ml 0.1N HCl.
Take 1 ml of aliquot of blank and Transfer the content in to 25 ml
diluted B standards into a 10 ml volumetric flask mix and make up the
polypropylene tube, add 2 ml of buffer volume to 25 ml with distilled water. For
solution and mix. Add 2 ml of azomethine analysis of B take 1 ml of the aliquot and
H reagent, mix and after 30 minutes read proceed as for the standard curve.
the absorbance at 420 nm on 3. Water analysis : Take suitable
spetrophotometer. With the help of quantity of water sample (containing 0.2
absorbance readings of standard solutions to 5.0 µg B) in porcelain dishes add 2 ml
of different concentration of B the Ca(OH)2 and proceed as described for soil
standard curve is drawn and a factor for extract. It is important to keep a definite
concentration of B for 1 absorbance is volume of aliquot i.e. 1 ml of either soil,
calculated which is utilized to calculate B plant or water in final step of B
in the soils, plant or water sample. determination.
Preparation of Extracts : References :
1. Soil extracts : The hot water Berger, K.C. and Truog, E. (1939). Boron
soluble extraction procedure of Berger determination in soils and plants. Ind.
and Truog (1939) is being used widely Eng. Chem. Anal. Ed. 11 : 540-545.
with slight modification of adding dilute Bighman, F.T. (1982). Boron p. 501-508. In
electrolyte (0.01 M CaCl2) instead of A.L. Page (ed.) Methods of Soil
water only. This provides clear, colourless Analysis. Part II Agron. Monogr. 9
extract which eliminates the need of ASA and SSSA, Mandison, W.I.
charcoal for decolourzation. Beside this a Jackson, M.L. (Ed.) (1958). Boron
determination for soil and plant
negative error, associated with B
tissues. In Soil Chemical Analysis
adsorption by charcoal, is also removed. page 370-387.
Place 20 g air dry soil in 250 ml John, M.K., Chuah, H.H. and Neufld, J.H.
low B flat bottom flasks and add 40 ml of (1975). Application of improved
0.01 M CaCl2 solution. Attach water azomethine H method for the
cooled reflux condenser to the flask. Heat determination of boron in soil and
the flasks for 5 minutes and then cool and plants. Anal. Lett. 8 : 559-568.
filter the suspension in plastic bottles.


Profile Studies of Deep Black Soils

G.P. Gupta
Deep black soils or Vertisoils occur shaped aggregates, diapir (mukara), and
globally under various parent materials gilgai. Shrink-swell phenomena are the
and environmental conditions (Table 1). dominant pedogenic processes in
Vertisols and are attributed to changes in
Table 1. Distribution of Vertisols and interparticle and intraparticle porosity
associated soils with changes in moisture content.
Juris- Location Area % of
Diction (m ha) Gross Definition of Vertisols :
Black
soils Taxonomically for defining
Continent Africa 105.0 38.7 Vertisols, there must be
Asia & far 70.3 25.9 1. A layer 25 cm or more thick with an
East (Mainly upper boundary within 100 cm of the
India) mineral soil surface, that has either
Australia 48.8 17.7
Latin 27.0 9.9
SLICKENSIDES or WEDGE
America SHAPED PEDS that have their long
North 10.0 3.7 axes tilted 10 to 60o from the
America horizontal; and
Near & 5.7 2.1 2. A weighted average of 30 % or more
Middle East
clay in fine earth fraction either
Europe 5.4 2.0
TOTAL 271.4 100
between the mineral soil surface and a
Country India 70.3 25.9 depth of 18 cm or in Ap horizon,
Australia 48.8 17.7 whichever is thicker and
Sudan 43.4 16.6 3. 30 % or more clay in fine earth
USA 18.1 6.7 fraction of all horizons between a
CHAD 15.5 5.7 depth of 18 cm and either a depth of
China 11.6 4.3
Others 64.5 23.7
50 cm or a densic, lithic or paralithic
( in parts) contact, a duripan, or a petrocalcic
TOTAL 271.4 100 horizon if shallower and
India MS 24.2 34.4 4. Cracks that open and close
MP 21.2 30.1 periodically.
GUJ. 4.9 7.0
AP 9.4 13.4
Vertisols are significant global
KTK. 5.8 8.2
TN 2.6 3.7
resources that serve as the lifeline in
RAJ. 1.1 1.6 subsistence agriculture due to their high
UP 1.1 1.6 productivity.
TOTAL 70.3 100 Efforts towards comprehension and
MP Vertisols 8.0 37.7 successful utilization are imperative for
Inceptisols 8.6 40.6 continued productivity and long term
Entisols 4.2 19.8
sustainability of these resources for
Alfisols 0.4 1.9
TOTAL 21.2 100 current and future civilizations.
They clayey soils that shrink and Morphology of a soil is best evaluated
swell extensively upon changing soil from the in situ examination of the soil
moisture conditions. Vertisols exhibit profile. A recently dug pit large enough
unique morphological properties such as for observation of a pedon is desirable.
the presence of slickensides, wedge- Old exposures such as road banks and


ditches are acceptable only for (ii) Weathering period must be

preliminary studies because extensive for the development of
morphological features often become solum with 2:1 expanding clays
altered after prolong exposure. (iii) Weathering environment should
Normally the size of profile pit is be such that no further weathering
kept 1.8 m long, 1.2 m wide and 1.8 m of 2:1 expanding clays takes place.
deep but for the study of black soils, the Even no inter-layering exists to
width of pit varies from place to place the extent the properties are
depending on its cyclic wave length of destroyed
puffs and shelves. It should be kept in (iv) Sequence of events should
mind that atleast half wave length continue like churning/mixing,
covering both, puff and shelve is development of argilli-
considered while exposing profile pit in pedoturbation, development of
order to study the pattern of cracks and slickensides and formation of
slickensides perfectly. wedge shaped structures
The profile examination begins Pedogenesis of Vertisols :
with a first approximation and marking of
soil horizon boundaries on the profile. 1. Separation of blocks :
Each horizon is then carefully observed Deep wide cracks separate the soil
and described. Horizon boundaries are into strong and massive prism like blocks
relocated as required by the detailed study in the upper part of the pedon that break
(Buol et al. 1998). Th description sheet into angular blocky peds of hard and firm
containing the columns of site and soil consistence.
characteristics is filled up by the profile (a) Cracking of soil : During dry season,
study group during pedon studies. the soil cracks to the surface due to
Vertisols are relatively shrinkage of 2:1 expanding clays that
homogneous in their morphology. may extend to a depth of 1 metre or
Although horizonation is not distinct yet a more.
few horizons above the parent material (b) Falling of surface soil material :
may be identified as self mulching surface While cracks are open, surface soil
(Ap), blocky subsurface (A12), material falls into them by several
slickensided horizon and wedge shaped mechanisms such as animal activity,
subsoil (Bss). wind or at the onset of rainy season by
The depth of these soils may vary water.
from shallow to very deep. Previously the 2. Hydration of clays : The clay
black soils were grouped as shallow (<30 hydrate and due to their high coefficient
cm) medium (30-100 cm) and deep (>100 of expansion and contraction, expand 3
cm) but lateron Sehgal (2008) modified dimensionally on wetting.
the dept of shallow soil as less than 50 (a) Expansion of clays : Cracks close on
cm. the surface but because of the extra
Requirement of Vertisols : material now present in the lower part
of the profile, a greater volume is
Main requirements of Vertisols are attained and the expanding material
the presence of high content of clay (>30 presses and slides the aggregates
%) and predominance of montmorillonite against each other, developing a
(2:1 expanding clay). Other important "lentil" angular blocky structure with
parameters for the development of sliekenside features on the ped
Vertisols are: surfaces.
(i) Parent material having basalt, (b) Shear stress development : The
argillaceous limestone, marine slipping occurs where shear strength
clays and shales is surpassed by shear stress acting


upon a soil mass. The shear stress is a Hallsworth and Beckman (1969)
major force caused by swelling and classified gilgai into 6 types i.e. normal or
develops when volume expansion round, melon hole, Lattice, Linear or
results during the wet cycle. wavy, tank or stony but lateron Paton
(c) Formation of slickensides : (1974) suggested only two types of gilgai
The slickensides, intersecting or close i.e. linear and circular (Nuram or
enough to intersect, also result in Pockmarked) each of which were grouped
wedge shaped structural aggregates, into 4 types.
the most characteristics feature of  type - Mound and depression equally
Vertisols which develop with their developed (No shelf present)
longitudinal axes inclined at 30 to 60°  type - Mound of much greater extent
from horizontal (Sehgal and than depression (No shelf present)
Bhattacharjee, 1988).  type - Depression of much greater
(d) Buckeling of land space : This extent than mound (No shelf present)
expansion buckles the land scape,  type - Mound, shelf and depression all
forming the micro relief called gilgai. present
The micro basins contain more 2. Size of cyclic pedons : Half cycle
organic matter than the micro ridges linear distance (HCLD) measures the
and probably it results from lateral dimension of a cyclic pedon. It
admixtures of subsurface material into may be small, medium or large i.e. below
micro ridge area and slight erosion of 1, 1 to 2 or above 2 to 3.5 metre,
organic rich fines from the ridges to respectively.
the basins. 3. Horizon sequence : In Vertisols,
3. Incomplete leaching : In most the horizon sequence has been suggested
shrink swell soils, the temperature being to be A1-Bss-BC-C where "ss" indicates
high, the potential evapotranspiration about the presence of slickensides.
suggesting incomplete leaching and 4. Thickness of horizon : Thickness
inducing the process of calcification in of A1 in Vertisols varies with the linear
these soils. frequencies of puffs and shelves of gilgai
Cyclic movement of soil material : micro relief.
Amongst several processes acting 5. Horizon boundary (Amplitude):
in the formation of Vertisols, the It is the difference between vertical distance
predominant process seems to be from the surface of pedon to the lower
haploidization i.e. mixing by argilli boundary of crest of cycle and the lowest
pedoturbation. The specific features of point of trough of cycle in same pedon. The
such soils are : amplitudes are grouped as low, medium of
1. Gilgai micro relief : The term high according to the vertical distance as
gilgai is an Australian aboriginal term below 25, 25 to 75 or above 75 cm,
meaning small water hole. respectively. Shape of apparent topography
Pedogenic micro topographical of the intermittent horizon is also graded as
features like puffs (microknolls) and tongued (vertical extent > horizontal
shelves (microbasins) develop that remain distance), wavy (vertical extent
intimately associated with one another approximating the horizontal distance) and
(Bhattacharjee et al. 1977), Columbe et smooth (vertical extent < horizontal
al. (1996) introduced a term "diapir" distances) as suggested by Bartelli (1971).
meaning a protusion of subjacent soil Age of Vertisols :
material which penetrates to the overlying It is difficult to assign the Vertisols
horizons and approaches or reaches the a place in the genetic scheme of soil
classification as there are greater
surface. If diapir and gilgai occur, the
differences of opinion whether they are old,
mound in gilgai is always developed over
young or remain in equilibrium with the
the diapir.
environment.


1. Views as Vertisols are old : The dominate. This interpretation suggests

end product of a development sequence that Vertisols are relatively young soils.
involves the soils whose B horizon has 3. View as Vertisols are in
become so clayey that shrink-swell cycles equilibrium : Vertisols remain in
developed and eventually "swallowed" equilibrium with their environment and
the A horizon. It is possible because high that the 2:1 expanding lattice clays are
content of fine clay and high fc/cc ratio stable and will persist, barring a climate
may be produced by lessivage on a large change. Vertisols then can be considered
scale. diagnostic of environments in which the
2. Views as Vertisols are young : parent material is basic and gives rise to
The fate of Vertisol may be to undergo the formation of 2:1 expanding lattice
alteration of 2:1 clays to non expanding silicates under the influence of wet dry
type of clay. The profile would then cease climate.
to churn and eluviation process would
Table 2 : Range in characteristics of Vertisols and Vertic Inceptisols
Horizon Soil colour Texture Structure Special features Width of
(10 YR) cracks (cm)
A. Typic Haplustert (10 YR - 2.5 YR)
Ap/A11 4/2, 3/3, 3/2, 3/1 C 1f/1m sbk 1c/2c pr-3c pr 2-5
A12 3/3, 3/2, 3/1 C 2m/2c abk 2c pr - 3c pr 2-5
Bss 3/3, 3,2, 3/1, 2/1 C 2m/3c abk Intersecting lickensides* 1-2
BC 4/4, 3/4 C 2m/2c abk ----do---- 0.5-1
C 5/4, 4/4, 4/3 c-gc 2msbk/ massive - -
B. Vertic Haplustept (10 YR - 7.5 YR)
Ap/A 5/2, 4/3, 4/2, 3/2 Cl gr/1m sbk 1c pr-2c pr 2-2.5
AB 4/3, 3/3 cl-c 1m/2m sbk ----do---- 2-2.5
B21 4/3, 3/3, 3/2 cl-c 2m sbk-3m/3c 2c pr - 3c pr or pressure 1.5
faces/abk slickensides
B22 6/3, 5/3, 4/4, 4/3 gscl-cl ----do---- - -
C 7/6, 6/3, 5/3, 4/4 gsl-gscl 1f sbk/ massive - -
*or parallelepipeds with long axes tilted from 35° to 55° from horizontal
References : Paton, T.R. (1974). Origin and terminology
for gilgai in Australia. Geoderma. 11 :
Bartelli, L.J. (1971). Soil taxonomy, 221-242.
correlation and Interpretation. Sehgal, J. (2008) Pedology – concepts and
Proceedings of Workshop organized by application, 2nd edition Kalyani
All India Soil and Land Use Survey, Publication, Ludhiana P. 416.
New Delhi. Sehgal, J., and Bhattacharjee, J.C. (1988).
Bhattacharjee, J.C., Landey, R.J. and Typical Vertisol of India and Iraq-their
Kalbande, A.R. (1977). A new approach characterization and classification.
in the study of Vertisol morphology. J. Pedologie. 38 : 67-95.
Indian Soc. Soil Sci. 25 : 221-232. Sehgal, J., Saxena, R.K. and Vadivelu, S.
Buol, S.W., Hole, F.D. , McCracken, R.J. and (1987). Field Manual for Soil Resource
Southard R.J. (1998). Soil Genesis and Mapping of different states. NBSS &
classification. Panima Publication LUP, Nagpur, pp. 73.
Corporation, New Delhi. Soil Survey Staff (1999). Keys to Soil
Coulombe, C.E., Wilding, L.P. and Dixon, Taxonomy. 9th Ed. CSC, SMSS, United
J.B. (1996). An Overview of Vertisols: States Department of Agriculture
Characteristics and Impact on Society. (USDA) Washington, D.C.
Adv. Agron. 57 : 289-375.


Food Preservation by Gamma Irradiation and its Importance

R.K. Thakur and G.D. Shrma
Gamma irradiation has been extensively tremendous potential as the world largest
used for food irradiation and sterilisation, food factory.
killing of fungus and micro organisms, It has been estimated that about 30-
sterilisation of medical accessories and 35% of fruit and vegetables of worth Rs
surgical equipments, high energy radiation 3000/- corers are perished every year. The
chemistry, seed irradiation and reasons for such losses are seasonal nature
semiconductor irradiation. Gamma of fruits and vegetables production. The
long distance between production and
Chamber can also be used in many other
consumption centers and also rising gap
research applications which require
between demand and supply. The hot and
irradiation of materials with ionizing humid climate in the country is also quite
radiations to varying doses. favorable for the growth of numerous
The radiation processing of food insects and micro organisms that destroy
involve the controlled application of energy stored crops and cause spoilage of food
from ionizing radiation such as gamma every year. The spoilage also occurs due to
rays, electrons and X rays for food chemical and physiological changes in
preservation. The gamma rays and X-rays stored foods. To preserve the food and food
are short wavelength radiation of products, technologies such as freezing
electromagnetic spectrum, which includes caning sun drying pickling fermentation
radiowaves, microwaves, infrared, visible, have been recommended by researchers but,
and a violet light. Radioisotopes such as each of these methods have its own merits
cobalt 60 and caesium-137 emit the gamma and limitation. The search for an alternative
rays, while machines using electricity newer economical methods to preserve food
generate electrons and X-rays. The gamma and causes least changes in sensory quality
rays and electrons are distinguished from have been under taken since long back, and
other form of radiation by their ionizing has been observed that radiation processing
ability. (That they are able to break of food is one of the latest method
chemical bond when absorbed by material). developed for food preservation.
The product of ionizing radiation may be
electrically charged ions) or neutral (free Irradiation by Gamma Chamber 5000 :
radicals). These there further react to cause Gamma Chamber 5000 is a
change in an irradiated material known as compact self shielded cobalt-60 gamma
the process of radiolysis. It is this reaction
irradiator providing an irradiation volume
that causes the death of micro- organism,
of approximately 5000cc. The material for
insect and parasites during food irradiation.
The conservation and preservation
irradiation is placed in an irradiation
of food is a prerequisite for food security. chamber located in the vertical drawer
It provides self-reliance to nation. The inside the lead flask. This drawer can be
Indian Food Industries contributes about moved up and down with the help of a
25-28% towards GDP. The food processing system of motorised drive which enables
sector provides 60-65% employment with a precise positioning of the irradiation
turnover of in US$ 36.1 billion of which chamber at the centre of the radiation
US$ 27.8 billion in organized sector, any field.
change or any stagnation in technology will Radiation field is provided by a set
inevitably have very large impact through of stationary cobalt-60 sources placed in a
out the economy. India is a potential cylindrical cage. The sources are doubly
producer of fruits and vegetables live stock encapsulated in corrosion resistant
and marine products. India has a stainless steel pencils and are tested in

accordance with international standards. Features :

Two access holes of 8 mm diameter are  Safe and self-shielded: The shielding
provided in the vertical drawer for provided is adequate to limit the
introduction of service sleeves for gases, radiation field on the external surface
thermocouple, etc. A mechanism for of the unit, well within the permissible
rotating/stirring samples during levels. No additional shielding is
irradiation is also incorporated. The lead required for its installation and use.
shield provided around the source is  Automatic control of irradiation
adequate to keep the external radiation time: Built-in timer provides accurate
field well within permissible limits. The control of irradiation time from 6
Gamma Chamber 5000 unit can be seconds onwards. The unit can also be
installed in a room measuring 4 metres x operated manually. Solid state
4 metres x 4 metres. programmable controls have been
provided. In the event of power failure
GAMMA CHAMBER 5000 battery backup displays the
programmes.
 Manual control of irradiation
temperature: It is possible to irradiate
samples at low or high temperature by
circulating liquid nitrogen or hot air.
These can be introduced through the
service sleeves provided in the vertical
drawer. The irradiation temperature is
sensed by a thermocouple and
displayed on the panel.
 Remote operation: An additional
table top control panel is provided for
remote operation in addition to the
normal one provided on the unit.
 Dose uniformity: Stationary source
pencils, symmetrically placed in a
cylindrical cage ensure good
uniformity of radiation field in the
sample chamber. In addition a
mechanism is also provided for
rotating/stirring samples during
irradiation.
 Easy loading and unloading of
samples: Sample chamber extends to a
convenient height for easy loading and
unloading of samples.
 Safety assurance: The design of Gamm
Chamber conforms to American National
Standards, ANSI-N433.1-1977 for safe
design and use of self-contained dry
source storage gamma irradiators
(Category I). It also meets the
requirements of type B(U) package for
safety in transport of radioactive materials
as per AERB code No.SC/TR-1, 1986 of
Atomic Energy Regulatory Board of
INDIA.


Applications : are made from a material including

photographic film, Perspex and cobalt
Gamma Chamber is a versatile
glasses. The poly vinyl chloride (PVC)
equipment for research studies in many
dosimeters are impregnated with a dye.
fields such as:
The Hydrogen chloride is released from
 Radiobiology
the PVC by irradiation and it produces a
 Preservation of tissue grafts
 Mutation breeding
qualitative or quantitative change in the
colour of the dye to indicate the dose
 Food preservation
received.
 Sterile male insect technique
 Biological and genetic effects of Dose distribution :
radiations The penetration of gamma
 Radiation chemistry radiation depends on the density of the
 Radiation effects on materials food as well as the energy of the ray. At a
 Radiation sterilization density of 1000 kg m-3 half of the rays are
 Modification of properties of absorbed in 11 cm. Halving the density
materials approximately double the depth of
penetration. The uniformity of dose
Food Preservation by Gamma distribution can be expressed as a ratio of
Radiation D max : D min. For sensitive food such as
The radiation processing of food is chicken the ratio should be as low as
carried out inside an irradiation chamber possible1.5.
shielded by 1.5 - 1.8 meter thick concrete Potential Applications of Gamma
walls. Food either pre-packed or in bulk Radiation :
placed in suitable containers is sent into
The radiation dose administrated to a
the irradiation chamber with the help of
food depends on the resistance of the
an automatic conveyor. The conveyor
organisms present and the objective of the
goes through a concrete wall labyrinth,
treatment. The maximum recommended
which prevents radiation from reaching
dose is 15 kGy, with average dose not
the work area and operator room. When
exceeding 100 kGy. Various application
the facility is not in use the radiation
of this technology are as under:
source is stored under 6 meter deep water.
The water shield does not allow radiation 1. Sterilisation ( or radappertisation) :
to escape in to the irradiation chamber, It is possible to sterilize meat and
thus permitting free access to personnel to other product, the dose required exceed
carry out plant maintenance. For treating the current limit of 10 kGy. A dose of 48
food, the source is brought to the kGy is needed for 12 D reduction of Cl.
irradiation position above the water level butulinum. High dose makes the product
after activation of all safety devices. The organoleptically un acceptable.
goods in aluminum carriers or tote boxes 2. Reduction of pathogens (radicidation) :
are mechanically positioned around the
source rack and are turned round their Food poisoning bacteria such as
own axis, so that contents are irradiated salmonella typhimurium are less resistant
on both the sides. The absorbed dose is to radiation than Cl. Botulinum, and doses
determined by the residence time of the of 3-10 kGy are sufficient for destruction.
carrier or tote box in irradiation position. 3. Prolonging shelf life (or radurisation) :
Measurement of radiation dose : Relatively low doses are needed to
Placing dosimeters at various destroy yeast, moulds and non-spore
positions in a tote box or carrier we can forming bacteria. This process is used to
check the absorbed dose. The dosimeters increase shelf life by an overall reduction
of vegetative cells.


Table 1: List of radiation processing facilities available in the world :

S. No. Country No. of Food Commodities
irradiators
1. Algeria 1 Potato
2. Argentina 1 Spices, spinach, coca powder
3. Bangladesh 1 Spices, onion, dried fish
4. Belgium 1 Spices, dehydrasted vegetables, deep frozen foods
5. Brazil 3 Spices, dehydrasted vegetables, fruits, vegetables, grain
6. Canada 1 Spices
7. Chile 1 Spices, dehydrasted vegetables, onion, potato, poultry meat
8. China 11 Spices and vegetable seasonings, Chinese sausage, garlic,
apple, potato, onion, dehydrated vegetables, sauses, rice,
tomatoes
9. Croatia 1 Spices, food ingredients, dried beef noodles
10. Czech. 1 Spices, dry food ingredients
Repulic
11. Cuba 1 Potato, onion, beans
12. Denmark 1 Spices
13. Finland 1 Spices
14. France 5 Spices, vegetable seasonings, herbs, poultry (frozen
boneless chicken, dried fruit, frozen frog legs, shrimp)
15. Hungary 1 Spices, onion, wine cork, enzyme
16. India 2 Spices, onion, potato
17. Indonesia 2 Spices, rice
18. Iran 1 Spices,
19. Israel 1 Spices, condiments, dry ingredients
20. Japan 1 Potato
21. Korea 1 Garlic powder, spices, condiments
Republic
22. Mexico 1 Spices, dry food ingredients
23. Netherland 1 Spices, frozen products, poultry dehydrated vegetables, egg
powder, packaging material
24. Norway 1 Spices
25. Poland 3 Not specified
26. Peru 1 Spices, food additives, animal feed
27. South 4 Spices, shelf-stable food, fruits
Africa
28. Thailand 1 Spices, fermented pork sausages, enzymes
29. Ukraine 1 Grain
30. UK 1 Spices
31. USA 10 Spices, poultry, fruits, vegetables
32. Vietnam 1 Onion
33. Yugosla 1 Spices
(Source : ICGFI, Food & Environmental Protection Section, Update, 1997)


4. Control of ripening : Plant Mutation Breeding by

Fruits and vegetables can be
irradiated to extend their shelf life about 2-3 Gamma Radiation
time when stored at 100C The ripening and Plant mutation breeding by
maturation of fruits are arrested by inhibition
radiation has been investigated for long
of hormone production and interupting the
biochemical process of cell division.
time in many countries. New mutant
5. Disinfestations : varieties give us useful gene resources for
Grain and tropical fruits may be the security of food resources, the
infested with insect and larvae, they reduces conservation of our ecosystem, and the
export potential. A low dose below 1 kGy is promotion of new industries. Using
effective for disifestation radiation technique (gamma-rays, X-rays
6. Inhibition of Sprouting: and EB) 128 varieties were developed in
The technology is effective in Japan. Many new species were developed
inhibiting sprouting of potatoes, A dose of for disease resistant crops, i.e. 55 species
about 150 Gy has been recommended. of rice, 10 of barley and 2 of wheat. Other
Similar doses are also effective in preventing species of beans, fruits including pears
sprouting of onion and garlic. resistant for black spot decease, grass,
Benefits and limitation of gamma
vegetables, etc, were also developed.
radiation processing:
Recently, a lot of fascinating new
Benefits : mutants are generated by ion beams. Ion
1. Radiation processing is a cold process beams can frequently cause large DNA
and therefore, unlike heat, it can be used alterations such as inversion, translocation
on agricultural commodities without and large deletion rather than point
changing their fresh-like character mutation, which result in producing
2. Radiation processing dose not alter characteristic mutants otherwise
significantly nutritional value, flavour, attainable. Ion-beam irradiation of
texture and appearance of food Arabidopsis seeds has produced the UV-
3. Radiation using Cobalt-60 cannot induce B-resistant, the frilled-petal, and other
any radioactivity in food and does not novel mutants. The features of ion beams
leave any harmful or toxic radioactive in the mutation induction seem 1) to
residues on foods as is the case with
induce mutants with high frequency, 2) to
chemical fumigants
4. Due to the highly penetrating nature of show a broad mutation spectrum, and
the radiation energy, it is a very effective therefore, 3) to produce novel mutants.
method. New mutants of chrysanthemum and
5. Prepackaged foods can be treated for carnation with complex and striped
hygienization and improving shelf-life flower-color, and new flower-shape have
6. The radiation processing facilities are been produced and commercialized.
environment friendly and are safe to Nuclear techniques, in contrast to
workers and public around. conventional breeding techniques, are
Limitations: widely applied in agriculture for
improving genetical diversity. Unlike
1. Radiation processing is a need based conventional breeding procedures which
technology and cannot be applied to all involve, the production of new genetic
kinds of foods
combinations from already existing
2. Radiation processing cannot make a bad
or spoiled food look good parental genes, nuclear technology causes
3. It cannot destroy already present exclusively new gene combinations with
pesticides and toxins in foods high mutation frequency. Basic tool of
4. Amenability of a particular food nuclear technology for crop improvement
commodity to radiation processing has to is the use of ionizing radiation which
be tested in a laboratory causes induced mutations in plants. These


mutations might be beneficial and have disintegration per second. The common
higher economical values. multiple is the megabecquerel (1 mCi =
37 MBq).
Measures of activity (A) :
Half –life: The time (t) taken for the
The number of disintegrations, or radioactivity of a sample to fall to half its
decay events, or nuclear transformations, initial value.
in a sample per unit time is its activity A. t1/2= 0.693 / k
Two common informal units are Electron volt (eV): energy of radiation
disintegrations per second and (usually as mega electron volts (MeV).
disintegrations per minute. leV=1.602x 10-19 J
Curie (Ci) : The US unit of activity is the Grays (Gy): Absorbed dose (where 1 Gy
curie (Ci). 3.7x1010 disintegrations per is the absorption of 1 J of energy per
second. Common multiples are the kilogram of food)
millicurie and microcurie. Previously rods (radiological unit) were
Becquerel (Bq) :The SI unit of activity is used. 1 rad = 10-2 J kg-1
the becquerel (Bq). One becquerel is 1


Plant Tissue Culture Techniques for Mass Propagation of Banana

Sunil Kumar and Sharad Tiwari
Biotechnology Centre, J.N.K.V.V., Jabalpur
steam under pressure as in a domestic

Ex 1. Aseptic culture techniques for pressure cooker. Most instruments/
establishment and maintenance of nutrient media are sterilized with the
cultures use of an autoclave. The standard
Principle : conditions for autoclaving have a
temperature of 121ºC and a pressure
Maintenance of aseptic environment:
of 15 psi (Pounds per square inch) for
All culture vessels, media and 15 minutes to achieve sterility. This
instruments used in handling tissues as figure is based on the conditions
well as the explants must be sterilized. necessary to kill thermophilic
The importance is to keep the air surface microorganisms. The time taken for
and floor free of dust. All operations are liquids to reach this temperature
carried out in laminar air-flow, a sterile depends on their volume. It may also
cabinet. Infection can be classified in depend on the thickness of the vessel.
three ways:
Precautions:
1. The air contains a large quantity of
suspended microorganisms in the 1. Excessive autoclaving should be
form of fungal and bacterial spores. avoided as it will degrade some
medium components, particularly
2. The plant tissue is covered with
sucrose and agar breakdown under
pathogens on its surface.
prolonged heating.
3. The human body (a skin, breathe etc)
2. At the bottom of the autoclave the
carries several microorganisms.
level of water should be verified.
4. In general, the methods of
elimination of these sources of 3. To ensure that the lid of the autoclave
infection can be grouped under is properly closed.
different categories of sterilization 4. To ensure that the air- exhaust is
procedures: functioning normally.
5. Preparation of sterile media, culture 5. Not to accelerate the reduction of
vessels and instruments (sterilization pressure after the required time of
is done in autoclave) autoclaving. If the temperature is not
6. Preparation of sterile plant growth reduced slowly, the media begin to
regulators stocks (by filter boil again. Also the medium in the
sterilization) containers might burst out from their
7. Aseptic working condition closures because of the fast and
forced release of pressure.
8. Explants (isolated tissues) are
sterilized using chemical sterilants, 6. Bottles, when being autoclaved,
e.g. HgCl2 and NaOCl. should not be tightly screwed and
their tops should be loose. After
Sterilization: It follows that all the autoclaving these bottles are kept in
articles used in the plant cell culture must the laminar air-flow and the tops of
be sterilized to kill the microorganisms these bottles are tightened on cooling.
that are present. B. Filter sterilization: Some growth
A. Steam or Wet sterilization regulators like amino acids and
(Autoclaving): This relies on the vitamins are heat labile and get
sterilization effect of super-heated destroyed on autoclaving with the rest


of the nutrient medium. Therefore, it Plant growth regulators (PGRs) at

is sterilized by filtration through a a very low concentration (0.1 to 100 μM)
sieve or a filtration assembly using regulate the initiation and development of
filter membranes of 0.22 μm to shoots and roots on explants on semisolid
0.45μm size. or in liquid medium cultures. The auxins
and cytokinins are the two most important
C. Laminar Airflow Cabinet: This is the
classes of PGRs used in tissue culture.
primary equipment used for aseptic
The relative effects of auxin and cytokinin
manipulation. This cabinet is used for
ratio determine the morphogenesis of
horizontal air-flow from the back to
cultured tissues.
the front, Air is drawn in electric fans
and passed through the coarse filter Materials: Amber bottles, Plastic beakers
and then through the fine bacterial (100 ml, 500 ml and 1000 ml), Measuring
filter (HEPA). HEPA or High cylinders (500 ml), Glass beakers (50 ml),
Efficiency Particulate Air Filter is an Disposable syringes (5 ml), Disposable
apparatus designed such that the air- syringe filter (0.22 μm), Autoclaved
flow through the working place flows eppendorf tubes (2 ml), Eppendorf stand,
in direct lines (i.e. laminar flow). Benzyl-aminopurine (BAP), Naphthalene
Before commencing any experiment it acetic acid (NAA)
is desirable to clean the working MS Nutrients Stock Solutions: Nutrient
surface with 70% alcohol. salts and vitamins are prepared as stock
solutions (20X or 200 X concentrations of
Ex 2. Preparation of stock solutions of
that required in the medium) as specified.
MS (Murashige & Skoog, 1962)
The stocks are stored at 4ºC. The desired
basal medium and plant growth
amount of concentrated stocks is mixed to
regulator stocks.
prepare 1 liter of medium.
Principle:
The basal medium is formulated so MS major salts mg/L 500 ml stock
medium (20X)
that it provides all of the compounds
needed for plant growth and development, 1. NH NO
4 3
1650 mg 16.5 gm
including certain compounds that can be
2. KNO 1900 mg 19 gm
made by an intact plant, but not by an 3
isolated piece of plant tissue. The tissue 3. Cacl .2H O 440 mg 4.4 gm
2 2
culture medium consists of 95% water,
macro- and micronutrients, vitamins, 4. MgSO .7H O 370 mg 3.7 gm
4 2
aminoacids, sugars. The nutrients in the
5. KH PO 170 mg 1.7 gm
media are used by the plant cells as building 2 4
blocks for the synthesis of organic

molecules, or as catalysts in enzymatic
MS minor salts mg/L 500 ml stock
reactions. The macronutrients are required medium (200X)
in millimolar (mM) quantities while
micronutrients are needed in much lower 1. H BO 6.2 mg 620 mg
3 3
(micromolar, μM) concentrations. Vitamins 2. MnSO .4H O 22.3 mg 2230 mg
are organic substances that are parts of 4 2
enzymes or cofactors for essential 3. ZnSO 4H O 8.6 mg 860 mg

4. 2
metabolic functions. Sugar is essential for
in vitro growth and development as most 4. KI 0.83 mg 83 mg
plant cultures are unable to photosynthesize 5. Na MoO 2H O 0.25 mg 25 mg
effectively for a variety of reasons. 2 4. 2
Murashige & Skoog (1962) medium (MS) 6. CoCl 6H O 0.025 mg 2.5 mg

2. 2
is the most suitable and commonly used
7. CuSO 5H O 0.025 mg 2.5 mg
basic tissue culture medium for plant 4. 2
regeneration.


MS Vitamins mg/L 500 ml identical to the plant from which it was

medium stock derived. The totipotency of the plant cells
(200X) and tissues is the basis for in vitro cloning
1. Thiamine (HCl) 0.1 mg 10 mg i.e., generation or multiplication of
genetically identical plants in in vitro
2. Niacine 0.5 mg 50 mg
culture.
3. Glycine 2.0 mg 200 mg Micropropagation is used
4. Pyrodoxine (HCl) 0.5 mg 50 mg commercially to asexually propagate
plants. Using micropropagation, millions
Iron, 500ml Stock (200X) of new plants can be derived from a single
 Dissolve 3.725gm of Na EDTA (Ethylene plant. This rapid multiplication allows
2
diamine tetra acetic acid, disodium salt) in breeders and growers to introduce new
250ml dH O. Dissolve 2.785gm of cultivars much earlier than they could by
2
FeSO .7H O in 250 ml ddH O using conventional propagation
4 2 2
 Boil Na EDTA solution and add to it, FeSO
techniques, such as cuttings.
2 4 Micropropagation also can be used to
solution gently by stirring.
establish and maintain virus-free plant
Plant Growth Regulator Stock: The stock. This is done by culturing the plant's
heat-labile plant growth regulators are apical meristem, which typically is not
filtered through a bacteria-proof virus-infected, even though the remainder
membrane (0.22 μm) and added to of the plant may be. Once new plants are
autoclaved medium after it has cooled developed from the apical meristem, they
enough (less than 60ºC). The stocks of can be maintained and sold as virus-free
plant growth regulators are prepared as plants.
mentioned below.
Micropropagation differs from all
Plant Nature Mol. Stock Soluble
in
other conventional propagation methods
Growth Wt. (1mM)
Regulator in that aseptic conditions are essential to
Benzyl Autoclavable 225.2 mg/ml 1N achieve success. The process of
aminopurine NaOH micropropagation can be divided into four
(BAP)
stages:
Naphtalene Heat labile 186.2 mg/ml Ethanol
acetic acid 1. Initiation stage: A piece of plant
(NAA)
Indole-3- Heat labile 203.2 mg/ml EtOH/
tissue (called an explant) is (a) cut
butyric 1N from the plant, (b) disinfested
NaOH
acid (IBA) (removal of surface contaminants),
The desired amount of plant and (c) placed on a medium. A
growth regulators is dissolved as above medium typically contains mineral
and the volume is raised with double salts, sucrose, and a solidifying agent
distilled water. The solutions are passed such as agar. The objective of this
through disposable syringe filter (0.22 stage is to achieve an aseptic culture.
μm). The stocks are stored at –20ºC. An aseptic culture is one without
contaminating bacteria or fungi.
Ex 3. Micropropagation of Banana 2. Multiplication stage: A growing
through shoot tip culture explant can be induced to produce
Principle: Plant cells and tissues are vegetative shoots by including a
totipotent in nature i.e., every individual cytokinin in the medium. A cytokinin
plant cell or tissue has the same genetic is a plant growth regulator that
makeup and capable of developing along promotes shoot formation from
a "programmed" pathway leading to the growing plant cells.
formation of an entire plant that is


3. Rooting or preplant stage: Growing 7. Add filter sterilized IBA (desired

shoots can be induced to produce amount, calculate) once the temperature
0
adventitious roots by including an of the medium cools down to 60 C.
auxin in the medium. Auxins are plant 8. Dispense the medium to sterilized
growth regulators that promote root Petridishes (25 ml medium/plate)
formation. For easily rooted plants, an
Preparation and inoculation of explant:
auxin is usually not necessary and
The 2-3 months old young, healthy
many commercial labs will skip this
suckers are selected for shoot tip explants.
step.
The adhering soil and dirt is removed.
4. Acclimatization: A growing, rooted Remove roots and prepare rhizome of 3-5
shoot can be removed from tissue cm with 2-4 inches of suckers, than wash
culture and placed in soil. When this thoroughly under tap water with Tween
is done, the humidity must be 20 for 10-15 min. There after dip the plant
gradually reduced over time because material in a solution of ascorbic acid 100
tissue-cultured plants are extremely mg/lit and citric acid 150 mg /lit for one
susceptible to wilting. hour. Sterilize the explants using 0.1%
Materials: Beakers, Measuring cylinders, HgCl2 for 7 min. subsequently the
Conical flasks, Cotton plugs, Myoinositol, explants are washed gently three times
Sucrose, BAP (1mM stock), Agar Agar, with sterile DDH2O (double distilled
Forceps, Blade Holder (No.3), Sterilzed water) in aseptic condition under laminar
blades (No.11), NAA (1 mM stock), flow. Shoot tips of 1-2 cm are excised and
Micropipettes, sterilized microtips, placed the rhizome pieces on MS
petridishes. medium, and incubated it at 25±2°C
temperature and 65-70 % RH in dark for
The shoot multiplication medium for 7-10 days, for plant regenerations.
Banana is MS basal + BAP (3 mg/l) +
NAA (0.5 mg/l) Multiplication of shoots: Shoot
multiplication is carried out on MS
Preparation of MS medium (1000 ml) medium containing 3-8mg/l of BAP
• MS Major (20X) 50 ml combined with 0.2-0.5mg/l of IBA. All
• MS Minor (200X) 5 ml cultures are maintained at temperature of
25±2°C under 16h photoperiod regime at
• MS Vitamin (200X) 5 ml
3000 lux and 70% RH.
• Iron (200X) 5 ml
Hardening of regenerated plantlets:
• Myoinositol 100 mg
Transfer the rooted plantlets to mixture of
• Sucrose 30 gm (3%) sand and FYM (3:1)/ Vermiculite to a
1. Add BAP at this stage (Calculate, how poly tunnel under poly house at 25-30°C
much to add) and 85-90% RH for 21-25 days. Plant the
2. Make final volume to 1000 ml by acclimatized plantlets in to polybags
double distilled water
containing sand, soil and FYM (1:1:1)
3. Set pH at 5.8
under net house conditions, after 3-4
4. Add agar agar 8 gm/L (0.8%), melt the
agar agar in microwave oven
weeks transfer polybags to open for field
0 plantation.
5. Sterilize the media at 15 psi/121 C for
15 minutes References :
6. After autoclaving, gently swirl the Murashige T & Skoog F (1962) A revised
medium to mix the agar. When the agar medium for rapid growth and
is completely dissolved and mixed, the bioassays with tobacco tissue
medium should appear clear and not cultures. Physiol. Plant 15: 473-497.
turbid.


Estimation of Microbial Biomass Carbon in Soil

R.K. Sahu
Microbial biomass carbon : chloroform free of ethanol and the bottom

whitish phase was collected.
It was estimated by chloroform 4. One set of soil samples were taken in
fumigation extraction method (Brookes et
crucible and fumigated with ethanol free
al., 1985).
chloroform in a vacuum desiccator. For
Principle: the purpose 20 ml ethanol free chloroform
Overnight fumigation of chloroform was taken in petridish and was placed
is done to kill al the organisms in soil inside the desiccator the bottom portion
samples; after which the amount of the inside the vacuum desiccators. It was
organic C in the sample can be measured by attached to the vacuum pump and the air
fumigation- extraction method. was evacuated until the chloroform starts
boiling to saturate the desiccators with
Fumigation extraction method:-
chloroform fumes. Then the vacuum
The microbial biomass constituents desiccator was kept in dark room for over
released by CHCl3 fumigation treatment can night.
be extracted directly through chemical 5. Next day the vacuum was released and
extractants and the readily oxidisable C chloroform was removed from the
contained in the extract can be measured desiccator.
through standard chemical procedures. 6. 10 g each of fumigated and non fumigated
Reagents soil samples were weighed in 150 ml
conical flask and 40 ml of K 2SO 4 (0.5 M)
1. Ethanol free chloroform was added to each flask. Samples were
2. Conc. H 2SO 4 shaken for 30 minutes on a rotary shaker.
3. 0.5M K 2SO 4: 43.563 g K 2SO 4 was 7. Both the samples were filtered with
dissolved in distilled water and diluted Whatman no. 42 filter paper, labelled and
with 500 ml. freezed until digestion.
4. 0.2 N K 2Cr 2O 7: 0.9808 g K 2Cr 2O 7 was 8. 10 ml of the filtrate was taken in 100 ml
dissolved in 100 ml of distilled water. conical flask and 2 M of K 2Cr2O 7 (0.2 N)
5. Orthophosphoric acid and 10 ml of con. H 2SO 4 was added to it.
6. 0.005 N Ferrous Ammonium Sulphate The contents of the flask were allowed to
(FAS): 3.92 g of ferrous ammonium cool for half an hour then 5 ml
sulphate was dissolved in 0.15 ml conc. orthophosphoric acid was added along
H 2SO 4 and then diluted to 2 liters by with 200 ml distilled water. Minimum two
distilled water. blanks were also run with 10 ml distilled
7. Ferroin/Diphenylamine indictor water.
Procedure 9. Few drops of diphenylamine indicator
1. 10g of soil (three sets of each soil were added and titrated against ferrous
sample) was weighed and kept in to 100 ammonium sulphate (0.2 M) till the colour
ml beakers each. changed from violet to green.
2. 8 ml of distilled water was added to both Calculation:
beakers and were incubated for seven Microbial carbon Fumigated C – Unfumigated C
days at 25 0C in an incubator. (ppm) = 0.44
3. 20 ml of chloroform was taken in a Reference:
separating funnel. It was washed two Brookes, P.C., Kragt, J.F., Powlson, D.S. and
times with conc. H 2SO 4 (with half of the Jenkinson D.S. (1985). Chloroform
volume of chloroform) and the acid fumigated and the release of soil
(bottom phase) was discarded. It was nitrogen. The effect of fumigation and
washed twice with the same volume of temperature. Soil Bio. Biochem., (17):
distilled water similarly to make the 831-835.




Application of Global Positioning System (GPS)

G.S. Tagore and PC Amule
Global Positioning Systems is the fact that  Satellite Health - Availability of Signal
the positioning signal is available to users in  Signal Strength - Quality of Signal
any position worldwide at any time. GPS is
the only fully-functional satellite navigation  Distance from the Reference Receiver
system (allow small electronic devices to  Radio Frequency (RF) Interference
determine their three-dimensional location
(Longitude, Latitude, and Altitude) in within  Loss of Radio Transmission from Base
a few meters using time signals transmitted
Applications of GPS in various fields
along a line of sight by radio from satellites. it
can be generated to a large community of Agriculture
likely to grow as there are multiple  Precision agriculture has developed due to
applications, ranging from surveying, GPS and GIS. GPS provides precision soil
mapping, and navigation to GIS data capture. sampling, data collection, and data
Structure analysis; enable localized variation of
chemical applications and planting density
The GPS consists of three parts:
to suit specific areas of the field.
 After soil testing farmers can create a
prescription map of their fields. This map
tells how much fertilizer needs to be
spread in certain areas rather than applying
a uniform application across the entire
field. This use of precision agriculture
saves money on chemicals and prevents
plants from being over-fertilized, all of
which benefit the environment. GPS
allows farmers to cut costs, save time, and
Factors that affect GPS become more efficient. It also can help
increase productivity and profitability.
There are a number of potential error sources
that affect either the GPS signal directly or  Ability to work through low visibility field
your ability to produce optimal results: conditions such as rain, dust, fog and
darkness increases productivity.
 Number of satellites - minimum number
required  Accurately monitored yield data enables
future site-specific field preparation.
 Multipath - reflection of GPS signals near
the antennae  Utilizing a handheld GPS unit, a farmer
can locate pumps, irrigation standpipes,
 Ionosphere - change in the travel time of wet and dry areas, cattle, and the location
the signal of fences that need mending
 Troposphere - change in the travel time of  Through a tractor-based GPS, a farmhand
the signal is told when and where to turn to begin
 Satellite Geometry - general distribution of tilling or harvesting each row of a field.
the satellites This can greatly reduce overlap, which on
a large farm saves hours of work.
 Satellite Geometry - or the distribution of
satellites in the sky - effects the  GPS unit allows farmers to record the
computation of your position. This is often location of weeds or insect infestations and
referred to as Position Dilution of On the go yield monitors record how much
Precision (PDOP). different parts of the field are producing.


Military Surveying and Mapping

The GPS was primarily developed for real
The high precision of GPS carrier phase
time military positioning. Military
measurements, together with appropriate
applications include airborne, marine, and
adjustment algorithms, provide an adequate
land navigation.
tool for a variety of tasks for surveying and
Navigation mapping. Fishermen can mark excellent
Navigation using GPS can save countless fishing spots and then go back to the same
hours in the field. Any feature, even if it is spot at a later date. Geodetic mapping and
under water, can be located up to one hundred other control surveys can be carried out
meters simply by scaling coordinates from a effectively using high-grade GPS equipment.
map, entering waypoints, and going directly
GPS are becoming very effective tools for
to the site.
GIS data capture. The GIS user community
Remote Sensing and GIS benefits from the use of GPS for locational
data capture in various GIS applications. GPS
It is also possible to integrate GPS positioning
allows farmers to cut costs, save time, and
into remote-sensing methods such as
become more efficient. It also can help
photogrammetry and aerial scanning,
increase productivity and profitability. The
magnetometry, and video technology. Using
GPS will soon be a part of the overall utility
DGPS or kinematic techniques, depending
of technology.
upon the accuracy required, real time or post-
processing will provide positions for the
sensor which can be projected to the ground,
instead of having ground control projected to
an image.


Appendix I
Conversion factors :
me. mg/eq.wt
ppm mg/l or µg/ml
1 ppm 2.25 kg/ha
10,000 ppm 1 per cent
1 mg/100 g 22.5 kg/ha
1 kg/ha 0.1 g /sq m
1 hectare 2.471 acres
1 acre 0.405 hectare
1 kg /ha 1.12 lbs/ acres
Mesh number 16/ mm
Normality Specific gravity X Purity X 10
(Liquid) Equivalent weight
Normality Wt of salt per liter X Purity
(Solid) Equivalent weight
Grams per liter Normality X Eq. Wt.
Organic matter Organic carbon X 1.724
Optical density 2 – log T (T= transmission)
Per cent by Wt. Grams of solute
100 g of solution
1 Angstrom (Å) 10-8cm or 10-10 m
10 (Å) 1 nanometer or 1 millimicrometer
Temperature conversion :
o
C/5 (oF-32) / 9
P X 2.29 P2O5
P2O5 0.44 X P
K X 1.20 K2O
K2O X 0.83 K
SX3 SO4
N X 1.12 NH3
Protein (%) Nitrogen (%) X 6.25
Velocity of light 3 X 10-10 cm / sec.
Velocity of sound 332 m / sec.


Appendix II
Percentage composition of manures and fertilizers
Material N P2O5 K2O Others

FYM 0.5-1.5 0.4-0.8 0.5-1.9 -
Compost (urban) 1.0-2.0 1.0 1.5 -
Green manure 0.5-0.7 0.1-0.2 0.8-1.6 -
Bone meal(steamed) 1.0-2.0 25-30 - -
Anhydrous ammonia 82 - - -
Ammonium chloride 24 - - -
Ammonium nitrate 33 - - -
Ammonium sulphate 20.6 - - 24 (S)
Ammonium phosphate 11 48 - -
Diammonium phosphate 21 53 - -
Calcium cynamide 20 - - -
Calcium nitrate 16 - - -
Sodium nitrate 16 - - -
Urea 46 - - -
Super phosphate - 16 - 12 (S)
Dicalcium phosphate - 23 - -
Tricalcium phosphate - 45 - -
Rock phosphate - 11-17 - -
Basic slag - 3.5-8 - -
Muriate of potash - - 60 -
Sulphate of potash - - 48 18 (S)
Zinc sulphate - - - 35 (Zn); 18 (S)
Zinc chelate - - - 14 (Zn)
Copper sulphate - - - 25 (Cu); 13 (S)
Ferrous sulphate - - - 19 (Fe); 19 (S)
Iron chelate - - - 5-9 (Fe)
Borax - - - 11 (B)
Sodium tetra borate - - - 14 (B)
Manganese sulphate - - - 26 (Mn)
Manganese chelate - - - 10-12 (Mn)
Calcium sulphate (Gypsum) - - - 18 (S); 33 (Ca)


Appendix III
Ready reckoner of fertilizer schedule at varying soil test values for different crops (kg ha-1)
Fertilizer Name Fertilizers Recommendations
Very Low Low Medium High Very High
PADDY (80:50:30)
Urea 260 215 175 130 85
Super Phosphate 470 390 310 235 155
Muriate of Potash 75 65 50 40 25
SOYBEAN (20:80:20)
Urea 65 55 45 35 20
WHEAT IRRIGATED (100:50:30)
Urea 325 270 220 165 110
WHEAT UNIRRIGATED (30:40:20)
Urea 80 65 30 15 -
Muriate of Potash 50 30 15 10 -
GRAM (30:60:30)
Urea 100 80 65 50 35
MOONG / URID / LENTIL / ARHAR (20:50:20)
Urea 65 55 45 35 20
PEA (35-75-30)
Urea 100 80 65 50 35
MUSTARD (60-30-20)
Urea 195 165 130 100 65
SUNFLOWER / SAFFLOWER (40:40:30)
Urea 130 110 90 65 45
MAIZE (120:80:60)
Urea 390 325 260 195 130
VEGETABLES (100:50:30)
Urea 325 270 220 165 110
JWAR (50:35:25)
Urea 190 160 120 90 60
Note: In wheat, paddy, mustard and maize the 50% of urea to be applied as basal doses. Rest 50%
may be applies in 2 to 3 split doses as top dressing.


Appendix IV
General recommended doses of micronutrient fertilizer
Micronutrient Material and doses for application

Zinc Zinc sulphate (25-50 kg ha-1) (25 kg 0.5% zinc sulphate +
ha-1 for light soils and 50 kg for 0.25%lime
heavy soils)
Iron Ferrous sulphate (25-50 kg ha-1) 1-2% ferrous sulphate + half
of lime
Copper Copper sulphate (10 kg ha-1) 0.1% copper sulphate +
0.05% lime
Manganese Manganese suphate (10 kg ha-1) 1% manganese sulphate +
0.25% lime
Boron Borax (10 kg ha-1) 0.2% borax
Appendix V
Rating of Nutrients:
Rating Organic carbon Available N Available P Available K

(%)
(kg ha-1)
Very Low < 0.3 150 <5 < 200
Low 0.3 – 0.5 150 – 250 5 – 10 200 – 250
Medium 0.5 – 0.75 250 – 400 10 – 20 250 – 400
High 0.75 –1.0 400 – 600 20 – 40 400 – 600
Very High > 1.0 > 600 > 40 > 600


Appendix VI
Performa for Soil Health Card
In-situ information
To be recorded while collection of sample Sample No………….
I. General Information :
Farmers Name
Age
Male/Female
Education
Address
II. Land details :

Name of the Area Survey Owned Leased Irrigated/ Soil
field (ha) No in/out rainfed type
III. Cropping details :
Survey Kharif Rabi

number
Crop/ Yield Crop/ Crop/ Yield Crop/
variety q/ha Variety variety q/ha Variety
proposed proposed


IV. Farmers Self Assessment-Score Card :

S. Parameters Ratings Details Year
No.
I II III
I Soil health
Biological activity (Deep Medium Shallow)
1 Earthworms Good Many holses and casts

>10 worms
Fair Few holes and casts >7-5
Poor Little sign of worm activity 0
- 3 worms
2 Birds Good Many birds follow plough /
following tractor during ploughing
plough
Fair Some birds
Poor Very few birds /sometimes
no birds at all
Plant growth and yield
3 Seed Good Seed come up quickly, and
germination even emergence
Fair Germination is uneven, takes
one or two days more for
emergence
Poor Germination is very poor
with high degree of
unevenness
4 Uniformity in Good Even stand in growth,
growth uniform green colour
Fair Slight variation in crop
height, moderate growth and
differences in colour
Poor Uneven stand, stunted
growth and stressed
5 Grain yield Good Good yield, and quality
Fair Average crop in the region,
Poor Poor crop in the area and
yield is very poor


Soil Analysis Report by laboratory Incharge/Technical Person
Name of laboratory :
Date of Sampling :
S. No. Soil properties Kharif Rabi

I II III I II III
1. Soil pH
2. EC (dsm-1)
3. Organic Carbon (%)
4. Available N kg/ha
5. Available P kg/ha
6. Available K kg/ha
7. Available S (mg kg-1)
8. Zinc (mg kg-1)
9. Iron (mg kg-1)
10. Manganese (mg kg-1)
11. Copper (mg kg-1)
Irrigation facility and water quality
Date of Sampling :
Irrigated/Un-irrigated/ Open well/ Kharif Rabi

Source of irrigation borewell /tank
I II III I II III
Average depth of ground

water
Annual average rainfall

(mm)
Normal onset of rainfall

(week/month)
Quality of irrigation Poor/medium/

water good


Recommendation:
Soil Testing Sample No. ………………..

Kharif Rabi
Details Manure/Fertilizer Manure/Fertilizer
Recommended Applied Recommended Applied
-1
FYM(tha )
Green manure(tha-1)
Nitrogen(kg ha-1)
 Urea
 DAP
 Complex
Phosphorus (kg ha-1)
 Super phosphate
 DAP
 Complex
Potash(kg ha-1)
Micronutrients(kg ha-1)
 Zinc sulphate
 Micronutrient
mixture
Biofertilizers
 Azosprillum
 Rhizobium
 Phospho bacteria
Other details :
Nearest Agriculture Department Office
Address and Phone number
Input dealers
Address and Phone number
Name of the bank
Account Number
Nearest KVK
Address and phone number
Soil Testing lab
Address and Phone Number

LabManual2012 PDF

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

LabManual2012 PDF

Uploaded by

Copyright:

Available Formats

INDEX

S. No. Title Page No.

01 Importance of Soil Testing, Collection of Soil Sample, its 1-4

02 Sampling, Processing and Storage of Plant Samples 5-7

03 Simultaneous Measurement of Bulk Density and Water 8-9

04 Estimation of Soil Moisture by Different Methods 10-12

05 Determination of Soil pH and Electrical Conductivity 13-14

06 Determination of Soil Organic Carbon 15

07 Determination of Available (Mineralization) Nitrogen in Soil 16-17

08 Determination of Total Nitrogen in Soil and Plant 18-19

09 Determination of Phosphorous in Soil and Plant 20-22

10 Determination of Potassium in Soil and Plant 23-24

11 Determination of Sulphur in Soil and Plant 25-26

12 Determination of Zn, Cu, Fe and Mn in Soils and Plants by 27-30

13 Estimation of Boron in Soils, Plants and Water 31-32

14 Profile Studies of Deep Black Soils 33-36

15 Food Preservation by Gamma Irradiation and its Importance 37-42

16 Plant Tissue Culture Techniques for Mass Propagation of 43-46

17 Estimation of Microbial Biomass Carbon in Soil 47

18 Application of Global Positioning System (GPS) 48

Importance of Soil Testing, Collection of Soil Sample, its Processing and

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 1

SELECTION OF SAMPLING POINTS

 At the sampling site, remove the surface

Parts of an Auger COLLECTION OF SOIL SAMPLE BY

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 3

An ideal time for soil sampling is just REFERENCES:-

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 4

Sampling, Processing and Storage of Plant Samples

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 5

samples by thorough washing, and The plant sample is diacid with

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 6

and let it cool. Then, add 10ml of glass If Ca and Mg is to be determined,

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 7

Simultaneous Measurement of Bulk Density and Water Content by

Recent advances is nuclear technology placed between a cesium source and

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 8

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 9

Estimation of Soil Moisture by Different Methods

Direct method (Gravimetric method) : Equipments and reagents :

Principle : neutron cloud developing when the Ra-Be

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 11

6. Cable - A 10 meter electric cable Divide readings by standard to

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 12

Determination of Soil pH and Electrical Conductivity

Determination of pH is actually a water and dilute to 1 litre. Add 1 ml of

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 13

Electrical Conductivity : Reagents :

Since conductivity is reciprocal of

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 14

Determination of Soil Organic Carbon

The majority of mineral surface soils Apparatus and Reagents :

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 15

Determination of Available (Mineralization) Nitrogen in Soil by Alkaline

Preparation of soil sample : Principle :

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 16

Procedure : Interpretation of results :

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 18

Determination of Total Nitrogen in Soil and Plant

Total nitrogen is estimated by the micro- The micro-Kjeldahl method consists of

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 18

2. Electronic balance liberated ammonia absorbed in 20 ml

JN Krishi Vishwa Vidyalaya, Jabalpur – 482004 (M.P.) 19