Printed AHS 2307 Experimental Design Notes

AHS 2307 EXPERIMENTAL DESIGN
EXPERIMENTAL DESIGNS
Most experiments try to verify certain hypothesis suggested by past experiences and observations. A plant breeders
wonders why farmers in Kisumu are slow in adopting the high yielding banana varieties developed at Mombassa.
He visits the banana growing around Kisumu and observes that the new varieties are much more healthy and look
more vigorous than the local ones in disease – free areas. However, in fields infested with Black Sigatoka, a very
destructive and prevalent disease of bananas in that region, the damage to the new varieties is more severe than to
the local varieties. He suspects and therefore hypothesizes that the new high yielding banana varieties developed in
Mombassa are more susceptible to black sigatoka than are the local ones.
At other instances, theoretical considerations may play a major role in arriving at a suggested hypothesis. For
example, it can be shown theoretically that a maize crop removes more nitrogen from the soil than is naturally
replenished during one growing season. Thus, we hypothesize that to maintain the yield potential of a maize farm,
supplementary nitrogen fertilizer must be added for every maize crop.
Once hypothesis is made then a design for the procedure to verify it must be made. This is the experimental
procedure, which usually consists of four parts:
1. Selecting the appropriate materials to test
2. Specifying the characters to measure
3. Selecting the procedure to measure these characters
4. Specifying the procedure to determine whether the measurements made support the hypothesis.
1 and 2 are easy for the subject matter to specify. In our banana example the test material would be the local and
the newly developed varieties. The characters to be measured would probably be disease infection and fruit
yield. For the example on maize the test would probably be one of the recommended maize varieties and the
fertilizer levels to be tested would cover the suspected range of nitrogen needed. As bananas, maize yield and
other agronomic characters would have to be measured.
Arriving at these decisions does not need knowledge in statistics. However the procedures on how the
measurements are to be made and how these measurements can be used to prove or disprove the populated
hypothesis depends upon the techniques developed by statisticians. For example, the precision in measuring a
character depends upon the experimental design, number of replications, and plot technique used. Furthermore,
the decision whether a certain observed difference is considered real or imaginary based on probability and
statistical reasoning.
Statistical tools have use for all kinds of research. A wide body of procedures has been developed for the
biological, physical and social sciences.
Choosing the correct statistical procedure for a give experiment is based on expertise in both statistics and the
experimental subject matter. It is necessary that agricultural researchers have the necessary statistical
background so that they can correctly choose the statistical technique most appropriate for their planned
experiments.
The Need For Statistical Evaluation

If one harvests two equal areas of groundnuts from a field the yield of nuts from the two areas whether they be
rows in length or halves of the entire field will rarely be the same. The weight of fruit from adjacent trees in an
orchard is rarely the same. Differences of this sort among crop units are due to genetic and environmental
differences beyond the experimenter’s control. They are not errors in the sense of being wrong, they represent
variability among experimental units call experimental error.
An experiment with a single replication provides a very poor measure of treatment effects, further since there
are no two experimental units treated alike, it provide no measure of experimental error. These difficulties are
overcome statistically by requiring the collection of experimental data that will allow an unbiased estimate of
treatment effects and the evaluation of treatment differences by tests of significance based on measuring
experimental error.
Treatments effects are estimated by applying treatments to at least two experimental units (usually more), and
averaging the results for each treatment.
Three important principles in all experimental designs essential to statistics are:
1. Replication: - means that a treatment is repeated two or more times. Its function is to provide an estimate of
experimental error and to provide a more precise measure of treatment effects. The number of replications
required in an experiment depends upon the magnitude of the differences you wish to detect and variability of
the data you are working with.
2. Randomization: -is the assignment of treatments to experimental units so that all units considered have
equal chance of receiving a treatment. It function to assure unbiased estimates of treatment means and
experimental error.
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3. Local control: - allows for certain restrictions on randomization to reduce experimental error. For
example, in the RCBD treatments are grouped in blocks that are expected to perform differently, allowing a
block effect to be removed from the total variation in the trial.
Characteristics of a Well Planned Experiment

a) Simplicity:- selection of treatments and the experimental arrangement should be simple, without
compromising the experiments objectives
b) Degree of precision: - an appropriate design and sufficient replication should ensure that the
probability is high that the experiment will be able to measure treatment differences with the degree of
precision required by the experimenter.
c) Absence of systematic error: -the experiment must be planned to ensure that experimental units
receiving one treatment, in no systematic way differ from those receiving another treatment so that an unbiased
estimate of each treatment effect can be obtained.
d) Range of validity of conclusions: -conclusions should have as wide a range of validity or possible. An
experiment replicated in time and space would increase the range of validity of the conclusions that could be
drawn from it. A factorial set of treatments is another way for increasing the range of validity of an
experiment. In a factorial experiment the effect of one factor are evaluated under varying levels of a second
factor.
e) Calculation of degree of uncertainty: - in any experiment, there is always some degree of uncertainty
as to the validity of the conclusions. The experiment should be designed so that it is possible to calculate the
probability of obtaining the observed results by chance alone
PROCEDURE FOR EXPERIMENTATION

Important steps are:
1. Definition of the problem. State the problem you are dealing with clearly and concisely. Once the
problem is understood you should be able to formulate questions, which, when answered will lead to solutions.
2. Statement of objectives: - in the form of questions to be answered, hypothesis to be tested, or the effects
to be estimated. When there is more tan one objective, list them in order of importance since this might have a
bearing on the experimental design.
3. Critical analysis of the problem and objectives. The reasonableness and utility of the aims of the
experiment should be carefully considered in light of the present status of knowledge concerning the problem.
Are the objectives of the experiment really important to a solution of the problem?
4. Selection of treatments. The success of the experiment rests on the careful selection of treatments, whose
evaluation will answer the questions posed.
5. Selection of experimental material. In selecting experimental material, consider the objectives of the
material and the population about which you wish to test for treatments.
6. Selection of experimental design. Choose the simplest design, which is likely to provide the precsion you
require.
7. Selection of the unit for observation and the number of replications. Decide on size and shape of the
field plots if you are dealing with field experiments with plants. Plot size and number of replications, should be
chosen to produce the required precision of treatment estimate.
8. Control of the effect of the adjacent units on each other. Accomplished through use of guard rows and by
randomization of treatments.
9. Consideration of data to be collected. Data collected should properly evaluate treatment effects in line
with the objectives of the experiment. In addition, consideration should be given to collection of data that will
explain why the treatments perform as they do.
10. Outlining statistical analysis and summarization of results. Write out the sources of variation and
associated degrees of freedom in the analysis of variance. Include the various F tests you may have planned.
Consider how the results might be used and prepare possible summary tables or graphs that will show the
effects you expect. Compare these expected results to the objectives of your experiments to see if the
experiment will give the answers you are looking for.
11. Conducting the experiment. Use procedures that are free from personal biases or favouritism. Make use
of the experimental design in collecting data, so that differences among individuals or differences associated
with order of collection can be removed from experimental error. Avoid fatigue in collecting data.
12. Analyzing data and interpreting results. All data should be analyzed as planned and the results interpreted
in the light of the experimental conditions, hypothesis tested, and the relation of the results to facts previously
established. Don’t jump to a conclusion, even though it is statistically significant if the conclusion appears out
of line with previously established facts. In this case, investigate further.
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13. Preparation of a complete, readable and correct report of the research. There is no such thing as a
negative result. If the null hypothesis is not rejected, it is positive evidence that there may be no real
differences among the treatments tested.
Most of the steps above are non statistical. Statistical analysis, however, is an important part of
experimentation. Statistical science helps the researcher design his experiment and objectively evaluate the
resulting numerical data. Practices the 3 ‘R’s of experimentation
a) Replicate – it is the only way you will be able to measure the validity of your conclusions from an
experiment.
b) Randomize – statistical analysis depends on the assignment of treatments to plots in a purely
objective, random manner.
c) Request help – ask for help when in doubt about how to design, execute or analyze an experiment.
NORMAL DISTRIBUTION
Most biological data when plotted in a frequency curve closely fit a mathematically defined curve called
No. of
plants
Height
the normal frequency curve.
Formula for describing a normal frequency curve is -
= N e-(y-)222
(2)
where f= frequency of occurrence of any variate given

y= any given variate
N= No. of variates in the population
Normal distribution only varies from one another with respect to their mean and/or standard deviation.
The mean determines the position of a curve on the horizontal axis. The standard deviation determines
the amount of spread or dispersion among the variates.
Means different, std

deviations equal Means equal, std
deviation
different
Arithmetic mean is the most common and actually the best measure of central tendency.  represents the
arithmetic mean of a population and y or x for the mean of a sample. Mu() is a parameter, a fixed value
that we seldom know and y is a statistic, a value that varies from sample to sample population. The
population mean definitions:-
μ = ( Σ Xi ) / N
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 (Sigma) tells you to sum all the values of Y2. The summation index I=1….N, says that the values of yi go
from the value of y1 to that of yN.
Since we seldom, y ever know the value of , we estimate it from a sample mean y, which is defined as:
Equation
R represents number of variates in the sample.

Often we wish to denote the difference between a variate (y) and a mean (Ÿ), such deviates are often
represented by a small letter y-Y- Y.
Grams per plant
Y y- Ÿ (Y- Ÿ)2
3 0 0
4 1 1
5 2 4
2 -1 1
1 -2 4
15 0 10
dry weight of 5 plants, Ÿ=3

the most common measure of dispersion is the standard deviation and its square, the variance. The
standard deviation of a population , and the variance 2, when estimated from a sample are symbolized
S and S2 respectively.
Pop. Variance: 2 =(y2-)2
N
N is number of variates in the population
The best estimate of 2 from a small sample (r<60) is S2 = (Yi-ÿ)2/ r-1
r is the number of variates in the population

(Yi -ÿ)2 is a sum of squares. The denominator r-1 is referred to as the degrees of freedom for the sample
usually one lass than the number of observations.
S2 = (Yi-ÿ)2/ r-1
= ((3-3)2 + (4-3)2+ (5-3)2 +(2-3)2+ (1-3)2 ) 5-1=2.5
S=2.5 =1.58 g/plant

A shortcut method for large samples is: (EQUATION)
(Yi)2/r is called the correction term, C. C= (Yi)2/r

r-1 is of the degrees of freedom, one less than the number of variates in the sample.
S2 = (32+42 + 52 + 22+12 -(3+4+5+2+22)/5)/5 - 1 =2.5
SAMPLING FROM A NORMAL DISTRIBUTION

After exposing a number of plots to a certain treatment, the treatment effect is estimated by calculating
the mean of the sample. Repetitions of the experiment (in effect drawing other samples) will produce a
series of different means. How well does a single mean represent the true treatment effect? This problem
is approached by calculating confidence limits, a range of values within which the true mean of the
treatment effect will fall unless we have drawn a very unusual sample.
The standard deviation of the population of means is called the standard error of a mean or just standard
error Ÿ. When Ÿ is estimated from a sample it symbol is SŸ.
The mathematical relationship between the variance of the parent population and the variance of a
population of means drawn from it is: 2Ÿ= 2/r r is the sample size on which the population of means is
based.
GRAPH
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Frequency distributions of populations of means, varying in sample size, generated by repeated sampling
from the same normally distributed population of plot grain yields. The distribution becomes narrower and
taller as sample size increases according to the relationship 2Ÿ= 2/r. Because of this relationship, 2Ÿ
can be estimated from only a single sample by S2Ÿ= S2/r. This relationship is used to calculate a
confidence interval about a sample mean. The relationship is also used repeatedly in the ANOVA when
estimating the variance per plot, S2, from a series of means assuming each mean is drawn from the same
population. In this case S2Ÿ is computed from the sample means as S2Ÿ= (Ÿi- Ÿ..)2/(n-1) and then
estimated S2by solving S2Ÿ=S2/r for S2= r S2Ÿ.
t DISTRIBUTION AND CONFIDENCE LIMITS

Looking at another repeated drawing of samples of a given size, say r = 5
For each sample compute Ÿ, S, SŸ and another statistic, t, where t=(Ÿ-)/SŸ

Organizing the large population of t values in a frequency distribution would yield a frequency curve like
the above one. There is a unique t distribution for each sample size. For a sample size of 5, 2.5% of the t
values will be equal to or greater than 2.776 and 2.55 will be equal to or less than -2.776. The two-tailed t
table shows probabilities for obtaining t values for the degrees of freedom for different sample sizes. E.g.
for df=10 the t value to be expected with a probability of 0.01 (1%) is 3.169.
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The foregoing figure shows the t distribution for a sample size of 5 compared to the normal distribution.
See that the t distribution is more variable than the normal distribution. The larger the samples size the
closer t approaches the normal distribution. When t values are based on samples containing 60 or more
variates they are approximately normally distributed as they closely estimate a normally distributed
statistic which is calculated as &=Ÿ- /Ÿ t and & only differ in the denominator. With small samples, SŸ
is quite variable from sample and therefore t is more variable than & whose denominator, Ÿ is a
constant. With larger samples, however SŸ is less variable and therefore t values more closely estimate &
values.
Confidence limits can be calculated for any random sample and  will fall within them with a specified
confidence. This is done by solving t = (Ÿ-)/SŸ for  and calling the resulting two values confidence
limits. CL = Ÿ  tSŸ if we wish to 95% confident that CL will contain  , we multiply SŸ by a tabular t value
depending on n -1 degrees of freedom and the 5% level of probability. For a sample where r = 5, SŸ is
multiplied by 2.776 e.g. if r = 5, Ÿ = 3, and S2 = 2.5; then SŸ = 2.5/5 = 0.707 and Cl.95 = 3  2.776
(0.707) = 4.96 to 1.04 gm/plant. With a confidence of 95% we can say that  lies in this range. We may
have drawn a sample whose Ÿ and/or S2 deviates efficiently from  and/or 2 so that CL.95 will not contain
. However the chance of drawing such a sample is only 5%.
Test of significance and statistical hypothesis- when comparing two or more treatment means an
assumption called the null hypothesis is applied which assumes that the treatments have no effects. The
probability is then tested that means as different as those of our samples would occur by chance alone if
the samples were indeed random samples from normally distributed populations with equal means and
variances. If our analysis leads to the conclusion that we would expect such mean differences quite
frequency by chance, we do not reject the null hypothesis and conclude that we have no good evidence
of a real treatment effect. If the analysis indicates that the observed differences would rarely occur is
random samples drawn from population with equal mean and variances, we reject the null hypothesis and
conclude that at least one treatment had a real effect. At least one of the means is said to be significantly
different from the others.
If the probability is 5% or less that the observed variation among means could occur by chance, we say
that the means are significantly different. If the probability is 1% or less that the observed variation among
means could be expected to occur by chance, the differences are said to be highly significant.
Computing t
J (y-ȳ) (y-ȳ) 2 T (y-ȳ) (y-ȳ) 2
25 0.1 0.01 27 0.36 0.1296
26 1.1 1.21 26 -0.64 0.4096
27 2.1 4.41 26 -0.64 0.4096
23 -1.9 3.61 27 0.36 0.1296
24 -0.9 0.81 27 0.36 0.1296
25 0.1 0.01 27 0.36 0.1296
25 0.1 0.01 26 0.64 0.4096
24 -0.9 0.81 27 0.36 0.1296
23 -1.9 3.61 26 0.64 0.4096
26 1.1 1.21 27 0.36 0.1296
26 1.1 1.21 27 0.36 0.1296
25 0.1 0.01 293 2.5456
25 0.1 0.01
324 16.93
Ȳ= 24.92308 df = 22 Ȳ= 26.63636

Pooled S2 = 16.93 + 2.539/12 + 10
= 0.885
S(y1-ȳ2)= √s2(n1 + n2)/(n1n2) = √0.885(24)/(143)
= 0.3847
t= (Ȳ1-Ȳ2)/S(y1-ȳ2)
=1.74/0.3847
=4.52**
Tabular t value at 5% = 2.074 1% = 2.819
The difference therefore, is highly significant
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F DISTRIBUTION
An F test is a ratio between two variances and is used to determine whether two independent estimates
of variance can be assumed to be estimates of the same variance. In the analysis of variance the F test is
used to test equality of means that is can it be reasonably assumed that the treatment means resulted
from sampling populations with equal means?
From a normal distribution draw 5 samples (n = 5), each having a specified number of variates e.g. (r =
9).
Calculate the means of these 5 samples. Estimate 2 by calculating S21….S25. Sum these estimates of 2
to obtain an average estimate: S2 = (S21 …+S25)/5
Estimate the variance of means (2Ÿ) from the means of the 5 samples:
S2Ÿ = (ÿ2 -Ÿ..)2/5-1. From S2Ÿ , again estimate 2 using the relationship S2 = rS2Ÿ , where r is the no. of
variates in each sample. Compute the variance ratio F
F = S21 (calculated from samples means)  S21 (calculated by pooling sample variances
The degrees of freedom for the numerator are n - 1=4 (where n is the number of samples) and for the
denominator n(r - 1)= 5(8)=40 (where r is the number of variates in each sample). Imagine that this
sampling procedure is repeated until all possible sets of samples have been drawn and recorded, the
frequencies curve has been plotted. The F value is 2.61 is the value beyond which 5% of the calculated
values fall. This is the value for the 5% level found in an F table for 4 and 40 degrees of freedom. In the
same way F values can be determined for other sample sizes, nos. of samples and for other levels of
probability (2.5%, 1%).
Since both variances in the F ratio are estimates of the same variance (2), the ratio will be close to one
unless an unusual set of samples has been drawn. The F distribution for the sample size we are
considering (n = 5, r = 9) will look like the graph in next page. The area under the curve represents the
frequency of obtaining any given F value. For any given draw of a set of samples of n = 5 and r = 9 the
chances of calculated F value being equal to or greater than 2.61 are 5%. Or, the chances are 95% that
any given draw of such a set of samples will produce an F value of less than 2.61. Note that the F test is
one -tailed. That is, we are not interested in the probability that F is equal to some value less than 1.
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ANALYSIS OF VARIANCE AND t TESTS
Before discussing complex experiments, the ANOVA procedure will be explained with a simple case of
two treatments, when each has been randomly assigned by 10 experimental units. Yields (100kg/ha) of
bean varieties 1 & 2 from plots to which the varieties were randomly assigned.
Varieties Replications Yi. Ÿi.

1 19 14 15 17 20 85 17 ÿi
2 23 19 19 21 18 100 20 ÿz
185 18.5 = Y..
To find out the variability called experimental error, we compute the variance of each sample (S 21 and
S22), assume they both estimate a common variance (2) and then estimate this common variance by
pooling the sample variances.
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Pooling S21 and S22 gives an estimate of 2 based on variability within the samples, which we will
designate as S2w:
S2w = S21 + S22 /2

= 6.5 + 4.0/2
= 5.25
Assuming the null hypothesis that these two samples are random samples drawn from the same
population and that therefore Ÿ1. and Ÿ2. both estimate the same population mean () we estimate the
variance of means (2ÿ) from the means of sample1 and 2.
We again estimate 2 using the relationship S2y = S2 /r and solve for S2 r is the no. of variates on which
each sample mean is based. We will designate this estimate of 2 as S2b.
We now have two estimates of 2: S2w based on the variability within each sample and S2b based on the
variability between the samples. Assuming the null hypothesis to be true we would expect S 2w and S2b to
be nearly alike since they both estimate the same variance (2). We can determine the probability of
obtaining divergent estimates of 2 by calculating an F ration and referring to a table of F values. For this
F ratio we always put the variance estimate from the sample (treatment) means (S 2b) as numerator and
the variance estimate from the individual variates as denominator. Thus, F= S2b / S2w.
If the two treatments (samples) come from populations having different means, S 2b will contain a
component reflecting this difference and will be larger than S2w. for our experiment, F= 22.5/5.25 = 4.29
The numerator, S2b is based on 1 degree of freedom, since there are two sample means. The
denominator, S2w is based on pooling the degrees of freedom within each sample. Each sample has 5
variates and therefore 4 df so the degrees of freedom for S2w are 4 + 4= 8
From an F table we look up the F values we would expect with a specified probability if the null
hypothesis is true and our sample means differ only by chance. For degrees of freedom 1 (numerator)
and 8 (denominator) we would expect an F value of 4.29 or larger with a probability of about 7%. In other
words, if the true mean difference is zero (1 - 2 = d =0), the chance of obtaining an estimate of d=
300kg per hectare is about 7%. Usually, we are not willing to sample that this event (which has a 7%
probability of occurrence), did not occur; therefore it will be unwise to reject the null hypothesis and
conclude that the mean of variety 1 is really different from the mean of variety 2. On the other hand, a
mean variety difference of 300kg per hectare if real represents a considerable economic gain. Therefore
we might decide to evaluate the two varieties in additional experiments.
ANOVA AND EXPERIMENTAL DESIGN
The principal difference among experimental designs is the way in which experimental units are grouped
or classified by treatments. In all designs, experimental units are classified by treatments, but in some
they are further classified into blocks, rows, main plots and so on. The analysis of variance uses the
means of these groupings, called sources of variations, to estimate mean squares. A mean square
estimating the dispersion among plot measurements resulting from random causes is also calculated - it
is called experimental error. In the absence of real differences resulting from means of treatments, blocks
or other sources of variation, these mean squares will on the average be equal. Only rarely will one mean
square deviate greatly from another by chance alone. When an F test indicates that the mean square
from one of the sources of variations is significantly greater than the mean square resulting from random
effects, we say there are real differences among the means of that particular source of variation. But
remember that there is always a definite chance that we will be wrong in such a conclusion. It is up to the
experimenter to select the odds at which it is believed there are real effects.
It is customary to describe results that are expected by chance 5% or less as significant and those
expected 1% or less as highly significant. When an experimenter uses the phrase "the treatments are
significantly different", what is really meant is that if the null hypothesis is true, the odds of obtaining such
mean treatment differences are only 5%. The experimenter is gambling that there was no such chance
occurrence in the experiment and that therefore, the significant result was due to a real treatment effect.
The ANOVA in its simplest form of two treatments randomly assigned to an equal number of experimental
units involves the following procedures:
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1. Calculating experimental error as the pooled variance of the two samples, for examples MSE = (S 21 +
S22)/2
2. Computing a mean square for treatments (MST) based on the null hypothesis that both sample
means estimate a common population, that is MST = r S 2y , where r is the number of variates in each
treatment mean.
3. Computing the F ratio: MST/MSE and comparing the calculated F value to a tabular F value to
indicate the probability of obtaining the calculated value by chance if the null hypothesis is true and
both sample means represent a common mean.
The statistical significance of a difference between two samples means can be tested by the F ratio in an
analysis of variance or by a t test. Both tests are statistical equivalent, t 2 = F. the analysis of variance and
the F test are usually easier to compute.
The means of differences between all possible pairs of sample means from two populations, X and Y, is
symbolized by d and is related to the means of parent population of means and individual variates as
follows:
d =µx - y = X - Y
The variance of means differences, d2 is estimated from two samples by S2d
S2d = S2x + S2dy = S2x/rx + S2y/ry and when S2x = S2y = S2 and rx = ry = r, S2d = 2 S2/r
Experimental designs arise from the way in which experimental units are grouped or classified.
The following is a step by step procedure for completing the ANOVA for the data given last week:
1. Outline the ANOVA table (below) by listing the sources of variation and degrees of freedom. There are
10 experimental units in the experiment and therefore 10 - 1 = 9 df in total.
Sources of Degrees of Sum of Mean Observed F Tabular F

variation freedom df squares SS square MS 10% 5%
Total 9 64.5
Varieties 1 22.5 22.5 4.29 3.46 5.32
error 8 42.0 5.25
These total degrees of freedom are then partitioned according to the experimental design. There are two
treatments, therefore 2 - 1 = 1 df. Degrees of freedom for error can always be obtained by subtraction, 9-
1=8, but also in this case by pooling degrees of freedom within each sample. There are 5 variates in each
sample, and therefore 5-1=4df; 4+4=8 df for error.
2. Compute the sum of squares for varieties (SSV) and the mean square for varieties (MSV)
SSV = Ÿ2i/r - Ÿ2../nr = 852 + 1002/5 - 1852/2(5)
= 3445.0 - 3422.5 = 22.5
MSV = SSV/(df)v = 22.5/1 = 22.5
Note that we use totals, not means, in computing SSV.

The following algebra illustrates why totals can be used in place of means to calculate sum of squares.
Based on the hypothesis that over two varieties are not different and both are samples from the same
population. we learned that a second estimate of 2 is obtained by S2b = r S2y, where v is the variance of
variety means and r is the number of replications in each variety mean. The mean square for treatments
(varieties in this case) is S2b, that is MSV = r S2y. Note that S2y = (ÿi -ÿ..)2/(n-1) and thus
MSV = r[(ÿi -ÿ..)2)/n-1]
since MSV = SSV/(n-1), SSV = r[(ÿi -ÿ..)2]. Previously, we saw that (ÿi -ÿ..)2 = ÿ2i -(ÿi.)2/n, so now we
can write,
ssv = r[ÿ2I - -(ÿi.)2/n]
Now we replace means with totals, noting that ÿI = Yi./r and that ÿi = Y../r and thus
SSV = r[ÿ2i./r2 - Y../r2(1/n)]
Carrying out the indicated multiplication, r[ ], gives SSV = (ÿ2i/r) - (Y2../rn) which is the formula previously
given. This formula involves some basic rules you should learn in order to compute sums of square from
totals:
a.) The first term Y2i./r tells you to sum the squares of the totals (variety totals in this case) and divide
by r, the number of varieties making up each total in the numerator.
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b.) The second term, Y2../nr = (Yii)2/nr is known as the correction term. It is the square of the sum of all
the varieties in the totals of the first term divided by the number of varieties in the sum (Y ..) being
squared.
c.) If all treatments do not have the number of replications, each total must be squared and divided by
the number of varieties it contains before summing, SST = Y2i/rI - Y2../ rI = (Y21/r1+….+ Y2n/rn) -
(Y2../r1+….+rn)
3. Compute the total sum of squares (SS). This 5. Calculate F ratio for varieties
step is done just before computing the sum of F= MST/MSE = 22.5/5.25 = 4.29
squares for error.
SS= Y2ii - Y2../nr 6. Look up the tabular F values for the levels of
= (192+142+….+182) - 1852/2(5) significance you wish to compare. Degrees of
=3487.0 - 3422.5 = 64.5 freedom pertaining to the numerator of the F ratio
and read cross the top of the table and degrees of
4. Compute the sum of squares and mean square freedom for the denominator are read down the left
for error (SSE and MSE) side.
SSE = SS - SST = 64.5 - 22.5 = 42.0
MSE = SSE/dfE = 42.0/8 = 5.25
A POPULATION OF MEAN DIFFERENCES
11
In addition to an F test we can also use a t test to evaluate the likely hood that two means are significantly
different. First we need to see how a population of mean differences is generated from a population of
normally distributed varieties. In particular we need to know parameters of this new population are related
to parameters of the parent population and to the population of means also generated in obtaining the
population of mean differences. If from two normally distributed populations, X 1, X2, …..Xn and Y1,
Y2……Yn, we draw all possible samples of a give size and calculate their means, we will have two
additional populations, X1, X2, …..Xm and Y1, Y2……Ym. now if we take all possible pairs of means and
subtract thus, X1- Y2, X1-Y2…X1-Ym, X2-Y1…X2- Ym…Xm-Ym, we will have a 5th population that of mean
differences.
The number of mean difference (Q) of this population will be much larger than the populations of X 1 and
Y1. If the number of means in these two populations both equal M, then Q = M 2.
The following relationships among the means and standard deviations of these populations can be
proven mathematically but will merely be stated here. The mean of the mean differences equals the
difference between the means of the sample means from populations X and Y, and this difference also
equals the difference between the mean of population X and population Y:
d = x-y=x-yy if the x=y, then d=0
The variance of the population of mean differences is  = (dI-d)2/Q and is equal to the sum of the
variances of the respective means. Thus, 2d=2x +2y. From two samples, 2d is estimated by S2d from
the variances of sample means: S2d= S2x + S2y. Since S2x= S2x/rx and S2y= S2y/ry,
S2d=( S2x/rx) + (S2y/ry).
The square root of the variance of mean differences is called the standard error of a difference. Often in
statistical analysis, one variance is estimated from another.
Important relationships among variances that you will use frequently are:
S2y = S2/r
S2d = S2x + S2y
S2d = S2x/rx + S2y/ry
And when rx = ry = r and S2x = S2y = S2, then S2d = 2S2/r
t TESTS FOR SIGNIFICANCE

The formula for t as applied to a population of a mean differences is t = (d - d)/Sd
For the data given two lectures ago we want to know the probability that sample 1 and 2 could have come
from populations having identical means (1 = 2). This is analogous to the discussion we have just had
where we referred to populations X and Y, only now we are calling them Y1 and Y2. The mean difference
of our sample means is d = 17 - 20 = 3 kg/hectare. The standard error of the difference is:
Sd = S2y1 + S2y2 =  S21/r1 + S22/r2 = 6.5/5 + 4/5 = 10.5/5 = 2.10 = 1.449
Assuming the null hypothesis that (d = 0), it is calculated as t = d - d/Sd = 3 -0/1.449 = 2.07
From the tables we can find the lowest value of t that has a 5% chance of occurring. Assuming that
21=22 we look up t based on the pooled degrees of freedom within the samples, in this case 4+4=8. The
expected t value for the 5% level of probability is 2.306 and thus our treatment difference is again judged
not significant. Note that t2=F, that is 2.072 = 4.285. Allowing for rounding errors this is equal to our
previously calculated F of 4.29.
The analysis of variance procedure and the calculation of an f value leads to the same conclusion as the t
test. The tests are equivalent while the analysis of variance procedure is usually easier to carry out. Also
it should be noted that at test is appropriate when 12. In this case the f test of the analysis of variance
is not valid. When 12 and r1 = r2 = r, the t value required for significance is for r-1 degrees of freedom.
In our example r=5 and the required t value at the 5% level would be the tabular value for 4df, or 2.776.
When r1r2, the required t value must be calculated as it is somewhere between the tabular t for r 1-1 and
r2-1 df.
12
COMPLETELY RANDOMIZED DESIGN & RANDOMIZED COMPUTE BLOCK DESIGN
Introduction
Single factor experiments are experiments in which only a single factor varies while all others are kept
constant. Examples are:
Yield test of several rice varieties
A fertilizer trial involving several rates of only one fertilizer element
A trial involving several plant densities
Complete block designs are fitted for experiments with small numbers of treatments e.g. a fertilizer trial
involving one variety and six rates of nitrogen fertilization.
Completely randomized design is most appropriate for experiments with homogeneous experimental
units.
Randomizing and laying out- randomization is accomplished by assigning treatment to experimental units
entirely at random. This can be done by using a random number table, by drawing cards, by throwing dice
etc. to show this, we shall discuss an experiment involving four treatments A, B, C, and D, each replicated
five times. The randomization and layout of plots are accomplished as follows:
1. Determine the total number of experiment plots. For our example, the total number of
experimental plots is 5*4=20
2. Assign a plot number to each experimental plot in any convenient manner, e.g. consecutively
from 1 to 20.
3. Assign treatments to the experimental plots by using one of the following schemes: by table of
random numbers
i. Locate a starting point in a table of random numbers by pointing a finger, with
your eyes closed, to any position in a page
ii. Using the starting point obtained in set (i), select n three digits numbers where
n is the total number of experimental plots (n=20 for our example). Three-digit
numbers are preferred since they are less likely to include ties. Indeed, any
higher digit numbers can be used. For example, starting at any point read
downward vertically to get 20 three-digit numbers.
andom Sequence Random Sequence Random Sequence Random Sequence
number number number number
937 1 749 6 918 11 549 16
149 2 180 7 722 12 957 17
908 3 951 8 243 13 157 18
361 4 018 9 494 14 571 19
953 5 427 10 704 15 226 20
4. Rank the 20 selected numbers from the smallest to largest

Random Sequence Rank Random Sequence Rank
number number
937 1 17 918 11 16
149 2 2 772 12 14
908 3 15 243 13 6
361 4 7 494 14 9
953 5 19 704 15 12
749 6 13 549 16 10
180 7 4 957 17 20
951 8 18 157 18 3
018 9 1 571 19 11
427 10 8 226 20 5
5. Assign treatments to plots. Use the rank as plot number and use the sequence in which the
random numbers occurred to refer to treatments. Thus in our example, assign treatment A (i.e.
the point treatment) to plot numbers 17, 2, 15, 7 and 19; treatment B to the plot numbers 13, 4,
18, 1 and 8; treatment C to plot numbers 16, 14, 6, 9 and 12; and treatment D to plot numbers 10,
13
20,3,11, and 5.
By drawing cards
1. Shuffle an ordinary pack of cards and draw 20 cards at random
2. Assign ranks to cards according to number and suit. The result as follows:
3. Follow the procedures stated in step 4 of the previous scheme. That is treatment A is
assigned to plot numbers 14, 7, 9, 15 and 5; treatment B t plot numbers 11, 2, 19, 13 and 18;
treatment C to plot numbers 16, 8, 10, 1 and 3; and treatment D to plot numbers 20, 6, 17,
12 and 4. Clearly, this procedure is not applicable when the numbers of experimental units
exceeds 52.
By drawing lots
1. Prepare 20 identical pieces of paper and label them with the letters A, B, C and D; each letter
appears fives times
2. Mix the 20 pieces of paper thoroughly and place them in a box (or a hat)
3. Pick out a piece of paper at random. If it is labelled B, the experimental plot number 1 will receive
treatment B
4. Without returning the drawn piece of paper to the box, draw a second piece at random. If it is labelled
A, the treatment A will be assigned to the experimental plot number 2.
5. Repeat the process until all 20 pieces of paper have been drawn
ANALYSIS OF VARIANCE
Completely randomized design with an equal number of replications
Analysis of variance is one of the most powerful and commonly used statistical tools in analyzing
experimental data. It allows for the subdivision of the total variability into several components. For a CRD,
there are two components of variation.
a.) That due to experiment
b.) That due to experimental error
The relative size of these two components provides the basis for determining whether the observed
differences among treatments are real or not. An investigation on the effects of foliar and granular
application of some selected insecticides for the control of the brown grasshoppers and stem borers will
provide our example. The experiment involved six treatments and a control, each replicated four times in
a CRD.
The steps involved in constructing the ANOVA are:
1. Group the data by treatments and calculate the treatment totals treatment means and grand total
2. Construct an outline of the ANOVA as follows:
Source of Degrees Sum of Mean Observed F Tabular F
variation of squares square 5% 1%
freedom
Treatment 6 5587175 931196 9.82** 2.57 3.81
Experiment
Error 21 1990237 94773
Total 27 7577412
cv = 15.1% **= significant at 1%
14
3. Determine the degrees of freedom (df) for each source of variation as follows:
Total df = total number of observations - 1
= 28 - 1
= 27
Treatment df = total number of treatments - 1
=7–1 =6
Error of which can be computed in either of two ways:
a.) As total df - treatment = 27 - 6 = 21 or
b.) As (total number of treatments)(total number of replications - 1) = (7)(4 -1)= 21
Treatment Treatment
Treatment Grain yields (kg/ha) Total mean
Malathion (1kg) 2537 2069 2104 1797 8507 2127
Malathion (2kg) 3366 2591 2211 2544 10712 2678
Thiodan 2536 2459 2827 2385 10207 2552
Diazinon-A 2387 2453 1556 2116 8512 2128
Diazinon-B 1997 1679 1649 1859 7184 1796
Karate 1796 1704 1904 1320 6724 1681
Control 1401 1516 1270 1077 5264 1316
Grand total and mean 57110 2040
4. Using X,s to designate individual measurements from each plot, t as the number of treatments and r
as the number of replications, calculate the sum of squares as follows:
Correction factor (c.f.) = (grand total)2/total number of observations

= (X)2/rt
= (57110)2/(4)(7)
= 116,484,004
Total SS = X2 - c.f
= [(235)2 + (2069)2 + …..+(1270)2 + (1077)2 ] - 116484004
= 7577412
Treatment SS = (treatment total)2/r – C.F
= (8507)2 + (10712)2 + …(5264)2/4- 116484004
= 5587175
Error SS = total SS – treatment SS
= 7577 412 – 5587175
= 1990237
5. Calculate the mean squares (MS) by dividing each SS by its corresponding degrees of freedom
Treatment Ms = treatment SS/t-1 = 5587175/6 = 931196 an
Error = error SS/t(r-1)
= 1990237/21
= 94773
6. Calculate the observed F value
F = treatment MS/error MS
= 931196/94773 = 9.82
7. Obtain the tabular F values using the treatment degrees of freedom and the error degrees of
freedom. The tabular F values for 6 and 21 degrees of freedom at 5% and 1% levels of
significance are 2.57 and 3.81 respectively.
8. Fill up the ANOVA table with values computed in steps 4 to 7.
9. Compare the observed F value with the tabular F values based on the following decisions:
a) If the observed F is as large as or larger than the tabular f at the 1% level, the differences
among treatment means are said to be highly significant. This result is indicated by placing
two asterisks on the observed F value
b) If the observed value is as large as or larger than the tabular F at the 5% but smaller than
the tabular f at the 1%, the differences among treatments are said to be significant. This is
indicated by placing one asterisk on the observed F value
c) If the observed F is smaller than the tabular F at the 5% level, the differences among
treatments are said to be non significant. This is indicated by placing ns on the observed F
value
15
In the present example, the observed F is larger than the tabular F at the 1% level. Hence, the
differences among treatments are said to be highly significant.
In other words chances are less than 1 in 100 that all observed differences among the seven treatments
could be due to chance.
Note that we can only conclude that there exists some differences between one or more pairs of the
treatment but cannot state which specific pair or pairs are causing the difference.
10. Compute the coefficient of variation
cv = (error MS/grand mean )*100 = (94773/2040)*100 = 15.1%
The cv value, which indicates the degree of precision in a particular experiment is generally placed
underneath the ANONA table.
The coefficient of variation is a good index of the reliability of an experiment. It expresses the
experimental error as a percentage of the mean and thus the higher its value, the lower is the
reliability of the experiment.
Coefficients of variation vary greatly with the type of experiment, the crop being tested and the
character being measured. An experienced researcher can make good judgment on the acceptability
of a particular cv for a given type of experiment.
ANALYSIS OF VARIANCE: Completely randomized design with an unequal number of

replications
CRD is one of the few designs tat allows you to use an unequal number of replications and is still
early to compute. This feature is most useful for studies where the experimental material precludes
using an unequal number of replications for all treatments. Some examples of this are:
a.) Experiments for comparing body length of different species of insect caught in an insect trap
b.) Experiments that may start with an equal number of replications but some experimental units
(plots) will probably be lost or destroyed during the experiment
When the number of replications per treatment differs, the computational procedures shown
previously are slightly modified. To illustrate this, modified procedure data on potato yield from the
performance of post-emergence herbicides will be used. The correction factor and the total SS are
computed as before:
C.F = (grand total)2/total number of observations
= (103301)2/40 = 266,777,415
Total SS = X2 - C.F = [(3187)2 + (4610)2 + …(1030)2]- 266777415 = 20,209,724
Rate (kg Time of Treatment Treatment
Treatment type a.i./ha) app. DAS Grain Yield (kg/ha) total mean
Brominal 2.0/0.25 21 3187 4610 3562 3217 14576 3644
Brominal 3.0/1.00 28 3390 2875 2775 9040 3013
Basagran 2.0/0.25 14 2797 3001 2505 3490 11793 2948
Basagran 2.0/0.50 14 2832 3193 3448 2255 11638 2910
Gramoxone 3.0/1.50 21 2233 2743 2727 7703 2568
Gramoxone 1.5 14 2952 2272 2470 7694 2565
2,4-D Amine 2.0/0.25 28 2858 2895 2458 1723 9934 2484
2,4-D Amine 3.0/.00 28 2308 2335 1975 6618 2206
Duron 2.0/0.50 28 2013 1788 2248 2115 8164 2041
Handweed twice __ 15 &35 3202 3060 2240 2690 11192 2798
control __ 1192 1652 1075 1030 4949 1237
a.i.= active ingredient DAS = days after seeding
Computation of the treatment SS however differs slightly from the method given earlier in that the divisor
of the term involving the square of treatment totals vary from one treatment to another. Each divisor is the
number of replications corresponding to the treatment concerned:
Treatment SS = i.T2i/rI- C.F

= [(14576)2/4 + (9040)2/3 + … +(4949)2/4] – 26677415 = 15,090,304
The error SS is computed from the previously given formula
Error SS = total SS - treatment SS
= 5,119,420
The mean squares and F values are then calculated using equations given before
16
Source of
Degrees Sum of Mean Observed F Tabular F
freedom
Treatment 10 15090304 1509030 8.55** 2.18 3.00
Error 29 5119420 176532
Total 39 20209724
c.v = 16.3% ** = significant at the 1%
RANDOMIZED COMPLETE BLOCK DESIGN

Ii one of the most widely used experimental designs in agricultural research. It is characterized by blocks
of equal size, each of which contains a complete set of all treatments. The chief advantage of this design
is that it reduces experimental error (through proper blocking) while retaining much of the flexibility and
simplicity of the completely randomized design.
Blocking- the primary purpose of blocking is to reduce as much as possible heterogeneity among plots
within each block. Proper blocking increases the differences among blocks while leaving plots within a
block more homogeneous. Knowing the fertility pattern in the experimental area is of great value in
achieving this. Rules to follow are:
1. When the fertility pattern of the experimental filed is known, orient the blocks so that soil differences
between blocks are maximized and those within blocks are minimized. For a field with a unidirectional
fertility gradient, long and narrow blocks should be used. Blocks should be oriented such that their
length is perpendicular to the direction of the fertility gradient. For example, for a field with a gradient
along the length of the filed, blocking should be made across the width of the field, cutting cross the
gradient.
2. When a fertility gradient occurs in two directions, with one direction perpendicular to the other, or
nearly so, other more appropriate designs e.g. Latin square design or covariance technique may be
used. If, however, a randomized complete block design must be used, most or less square blocks are
recommended.
3. When the fertility pattern is not known or when fertile areas occur in unpredictable spots, blocks
should be as square as possible.
Except for the different treatment assigned, plots within each block should be manage as uniformly as
possible. Data collection and all cultural and management practices, aside from the treatments being
studied, should be made at the same time and as uniformly as possible on all plots un each block. For
example, if application of insecticides or harvesting of a field experiment must be done over several days,
all plots in a block should be applied or harvested in the same day. Also, if more than one research
worker makes observation on the experimental area. If there is any likelihood that observations made on
the same plot would differ with the individual, then different persons should be assigned to different
blocks, i.e. one person should make the observations for all plots in a block.
Randomization and layout

Divide the experimental area into as many blocks as the number of replications. Each block is further sub-
divided into plots that correspond to the number of treatments. Since the randomized complete block
design specifies that all treatments must appear once in each replication, randomization is done
separately for each block. Randomization procedures similar to those described earlier can be used to
assign treatments to plots in each block.
Let us take an experiment where we have six treatments (A, B, C, D, E and F) to be replicated four times.
If the table of random numbers is used, we first select at random six three-digit numbers.
Number Sequence Then rank the Number Sequence Rank Number Sequence Rank
selected numbers
918 1 from the smallest to
918 1 6 918 1 6
772 2 the largest 772 2 5 772 2 5
243 3 243 3 1 243 3 1
494 4 494 4 2 494 4 2
704 5 704 5 4 704 5 4
549 6 549 6 3 549 6 3
Third, use the sequence in which the random numbers occurred as the treatment number and use the
rank as the plot number in the block. Thus, assign treatment A to plot 6, treatment B to plot 5, treatment
to plot 1, treatment D to plot 2, treatment E to plot 4 and treatment A to plot treatment F to plot 3.
17
Plot number Block 1
1 treatment C
2 treatment D
3 treatment F
4 treatment E
5 treatment B
6 treatment A
Fourth, repeat the first three steps for Block II then for Block III and finally for Block IV.
Yield of bean variety Roscoco - 4 replications:
Treatment Treatment
Treatment (kg/ha)Block I Block II Block III Block IV total mean
25 5113 5398 5307 4678 20496 5124
50 5346 5952 4719 4264 20281 5070
75 5272 5713 5483 4749 21217 5304
100 5164 4831 4986 4410 19391 4848
125 4804 4848 4432 4748 18832 4708
150 5254 4542 4919 4098 18813 4703
Totals 30,953 31,284 29,846 26,947 119,030 4,960
a.) RCB layout
Block I Block II Block IIIBlock IV
C A F E
D E D C
F F C D
E C A A
B D B F
A B E B
A hypothetical layout of CRD is also given to emphasize the differences in layouts. Note that in a RCBD,
randomization is restricted so that all treatments must appear in the same block, unlike the CRD, which
has no restriction, whatsoever.
18
ANOVA
Group the data by treatments and blocks and calculate 1. Determine the degrees of freedom (df) for each
treatment totals, block totals and treatment means (as source of variation as follows:
shown in data table).
Outline the analysis of variance as follows: Total df = total number of observations - 1 = 24 -1 = 23
Source of Degrees of Sum of Mean Tabular F Block df = total number of blocks - 1 = 4 - 1 = 3

variation freedom squares square Observed F 5% 1% Treatment df = total number of treatments - 1 = 6 - 1 = 5
Treatment
Error Error df = total df - block df - treatment df = 23 - 3 - 5 =
15
Total
Or = (block df)(treatment df) = (3)(5)= 15
Note that in addition to the two sources of variation for
CRD, a third component, block is also separable for
this design.
2. Calculate the various sums of squares (SS) 3. Divide each sum of square (SS) by its corresponding
degrees of freedom to obtain the mean square:
C.F = (grand total)2/(total number of observations)
Block MS = block SS/r – 1 = 1944361/3 = 648120
= (119030)2/24 = 590,339,204
Treatment MS = treatment SS/t – 1 = 1198331/5
Total SS = X2 - C.F
= 239666
= [(5113)2 + (5346)2 + …. + (4098)2 - C.f
Error MS = error SS/(r - 1)(t - 1) = 1658376/15
= 595,140,272 - 590,339,204
= 110558
= 4,801,068
4. Compute the observed F value for testing the
Block SS = (block total)2/t - C.f treatment effects:
=(30953)2 + (31284)2 + (29846)2 + F = treatment MS/error MS = 239666/110558 = 2.17

(26947)2)/6- C.F.
5. Obtain the tabular f value for 5 and 15 degrees of
= 592,283,565 - 590,339, = 1,944,361 freedom. The tabular F value for 5% and 15 levels of
significance are 2.90 and 4.56 respectively. Since
Treatment SS = (treatment total)2/r - C.F. the observed f value is smaller than the tabular F
value at 55 we conclude that the experiment failed to
= (20496)2 +…. + (18813)2 )/4- C.F. show any significant difference among the six rates
of seeding.
= 591,537,535 - 590,339, = 1,198,331
6. Compute the coefficient of variation:
Error SS = total SS - block SS - treatment SS
cv = error Ms/grand mean *100
= 4801068 - 1944361 - 1198331
= 110558/4960 *100 =6.7%
= 1658376
19
LATIN SQUARE DESIGN
The most important feature of the Latin square design is its capacity to block an area along two directions instead of
only one as with the RCBD. The two-directional blocking, referred to as rows and columns, is done by arranging the
treatments so that every treatment occurs only once in each row and once in each column. This allows us to measure
and remove from the experimental error the differences among rows as well as among column. Thus, the Latin
Square design is especially adapted for use in an experimental site where there are two fertility gradients running
perpendicular to each other. The stratification into rows and columns, while useful in removing variability along two
directions, also becomes a major restriction, requiring that the number of replications must equal the number of
treatments. This restriction is very significant because for a large number of treatments, the design becomes
impractical. On the other hand, with fewer than four treatments, the number of degrees of freedom associated with
experimental error becomes too small. Thus, the Latin square design is is generally used only for experiments with
four to eight treatments. Because of such inflexibility and limitation, the Latin square design has not been widely
used in agricultural experimentation despite its potential for controlling experimental error.
Randomization and layout

The layout for Latin square design is set up by selecting a sample Latin square plan from the table followed by the
randomization among rows and among columns. For an example, lets us consider an experiment involving five
treatments A, B, C, D and E.
The randomization procedure is as follows:
Step 1: take the 5*5 Latin square plan shown in the table.
A B C D E
B A E C D
C D A E B
D E B A C
E C D B A
Step 2: Randomize the row arrangement as follows:

a) Select at random five three-digit numbers from the tables e.g. 628, 846, 475, 902, and 452
b) Rank these numbers from lowest to the highest
Random number Selection sequence Rank

628 1 3
846 2 4
475 3 2
902 4 5
452 5 1
c) Use these ranks as the row number of the selected plan and the sequence as the row number of the
rearranged plan. That is, the third row of the selected plan becomes the 1 st row; the fourth row becomes the
second and so on. Thus, the new plan after row re-arrangement is
C D A E B
D E B A C
B A E C D
E C D B A
A B C D E
Step 3: randomize the columns using the same procedure as in step 2. For example, five random numbers were again
chosen from the table and ranked as follows:
Random number Selection sequence Rank
792 1 4
032 2 1
947 3 5
293 4 3
196 5 2
20
The rank row represents the column number of the plan of step 2 and the sequence that the numbers occurred in
represents the column number of the newly rearranged plan. That is, the fourth column of the plan of the step 2
becomes the 1st column becomes the second and so on. Thus, the resulting plan is:
E C B A D
A D C B E
C B D E A
B E A D C
D A E C B
This plan becomes the final layout of the experiment.
Analysis of Variance: Grain yield of 3 maize hybrids and a check variety

Grain yield (bags/ha)
Row Col 1 Col 2 Col 3 Col 4 Row total
1 32.8 (B) 24.2 (D) 28.5 (C) 26.9 (A) 112.4
2 29.5 (C) 23.7 (A) 28.0 (D) 25.8 (B) 107
3 33.4 (A) 14.2 (C) 33.3 (B) 23.6 (D) 104.5
4 31.3 (D) 25.8 (B) 33.1 (A) 13.2 (C) 103.4
Column total 127 87.9 122.9 89.5
Grand total 427.3
The previous data is on maize yield of 3 hybrids (A, B and D) and a check variety, conducted in Latin square design
experiment with 4 replications.
The step in the ANOVA procedure are:
Step 1: Arrange the data by row and column as done above. For each observation, the particular treatment involved
must be specified.
Step 2: Compute treatment totals and treatment means as follows:
Yield (bags/ha)
Treatment Total Mean
A 117.1 29.3
B 117.7 29.4
C 85.4 21.4
D 107.1 26.8
Step 3: outline the analysis of variance:

freedom
Row
Column
Treatment
Error
Total
Note that there are four sources of variation for the Latin square design compared with three for the RCBD and with
two for the TCBD.
Step 4: Determine the degrees of freedom (d.f) for each source of variation as follows:
Total d.f = total number of observations – 1 = 16 – 1 = 15
Row d.f = column d.f = treatment d.f = total number of treatments – 1,
Error d.f = total d.f – row d.f – column d.f – treatment d.f = 15-3-3-3 = 6 or
= (total numbet of treatments –1)(total number of treatments-2) = (4-1)(4-2)=6
Step 5: Compute the various sums squares (SS):
21
C.F = (grand total)2/total number of observations
= (427.3) 2/16 = 11411.581
Total SS = x2– C.F

= [(32.8)2 +(29.5)2 + … + (13.2)2] – C.F
= 565.569
Row SS = (row total)2/t – C.F

= [(112.4)2 +(107.0)2 + (104.5)2 + (103.4)2]/4 – C.F
= 12.062
Column SS = (column total)2/t – C.F
= [(1270.)2 +(87.9)2 + (122.9)2 + (89.5)2]/4– C.F
= 330.837
Treatment SS = (treatment total)2/t – C.F
= [(117.1)2 +(117.7)2 + (85.4)2 + (107.1)2]/4 – C.F
= 170.737
Error SS = total SS – row SS – column SS – treatment SS

= 565.569 – 12.062 – 330.937 – 170.737
= 51.833
Step 6: compute the mean square for each source of variation by dividing the sum of squares by its corresponding
degrees of freedom.
Step 7: Compute the observed F-values by dividing each mean square by the error mean square as follows;
F for row = 4.021/8.639 <1, F for column = 110.312/8.639 = 12.77 and F for treatment = 59.912/8.639 = 6.59
Step 8: Compare each of the observed F-values with the appropriate tabular values and declare the test for the
particular effect significant or not significant following the rules given earlier.
Step 9: Enter all values so far computed in the analysis of variance table. The final results are shown below.
Significant differences in the mean yields of the four maize varieties tested were obtained. To fin out whether every
of the three hybrids gave significantly higher yield than the check or whether there are any significant differences
among the three hybrids etc test procedures that compare among treatment means may be used. These will be
discussed later.
Analysis of variance on maize yield data.
freedom
Row 3 12.062 4.021 <1 ns 4.76 9.78
Column 3 330.937 110.312 12.77** 4.76 9.78
Treatment 3 170.737 59.912 6.59* 4.76 9.78
Error 6 51.833 8.639
Total 15 565.569
cv=11.0%, **=significant at 1% level, *=significant at 5% level, ns=not significant
Row and Column Blocking

The test for column differences above showed a highly significant result while that for row differences
gave a non-significant result. This indicates that the grouping of experimental plots into rows was not as
successful in reducing the experimental error as was the grouping by column. This would suggest that a
randomized complete block design with columns as blocks would have done similarly well
FACTORIAL EXPERIMENTS
Characterized by treatments composed of all possible combinations of levels in each of two or more
factors. A simple example is one with two factors each at two levels, e.g. the response of two potato
varieties to two levels of nitrogen. The treatments for such an experiment might be:
Variety A at 0kg N/ha
Variety B at 0kg N/ha
Variety A at 60kg N/ha
Variety B at 60kg N/ha
The term ‘factorial’ merely specifies how the treatments are formed. It does not in any way describe how
the treatments are to be arranged in the experimental plots as any design should. A factorial is not a
design. A factorial experiment still needs to be fitted to a particular design. For example, if the potato –
22
nitrogen were fitted into a randomized complete block design, then the correct description would be a
factorial experiment in the RCBD.
Factorial experiments have a major advantage over simple factor experiments in having versatility to
evaluate response to simultaneous changes in several factors.
Interaction: when two variable factors are being evaluated in a simple experiment, the following can
happen:-
Either the change in one factor does not influence the relative effects of the other factor (the two factors
do not interact with each other)
The relative effects of one factor depend upon the level of another factor (one factor interacts with the
other).
The size of this interaction can be measured only if the factor are tested together in a factorial
experiment. The data below is for the previously described experiment involving two varieties of potato
and two levels of nitrogen. Potato yield (kg/ha):
Variety 0kg N/ha 60kg N/ha Average
(a) no interaction
A 10 30 20
B 20 40 30
Average 15 35
(b) interaction present
A 10 30 20
B 20 20 20
Average 15 25
Hypothetical examples of no interaction between variety and nitrogen effects and the presence of an
interaction.
When there is no interaction, the relative yield of one factor (e.g. variety) remains the same regardless of
the level of the other factor (e.g. nitrogen). Thus in the previous example the difference between the two
varieties remains the same at both 0N and 60N. that is, the yield of variety B relative to variety A is 10
units higher at both fertilizer levels. Similarly, the application of 60kg N/ha increased yield of both varieties
A and B by 20 units.
(b) shows the case where varieties interact with fertilizers. Variety B is better yielding at 0N but variety A
is better at 60N. in other words, the relative performance of the two varieties at the two levels of nitrogen
are not similar. Likewise, while the application of 60kg N/ha increased the yield of variety A by 10 units, it
did not change the yield of variety B. when there is an interaction between two factors, two single-factor
experiments (one on fertilizers and another on varieties) conducted separately would not be revealed as
complete information as that obtained from a factorial experiment involving the two factors. If the two
varieties were compared in a single-factor experiment the researcher would have used only one fertilizer
level. But the decision as to which of the two varieties is better depends upon whether 0 or 60kg N/ha
was used. This illustrates how limited and even at times misleading, the information from a single-factor
experiment can be. When interaction is present, a factorial experiment would not only give a broader
basis for comparing the varieties but would also give additional information concerning the desirable
fertilizer rate for each of the two varieties. It is thus advisable that a factorial experiment, instead of
several single-factor experiments, be conducted whenever possible.
An illustration of three types of relationship between two factors at two levels each:
(a) shows no interaction, (b) shows a mild interaction where rankings do not change in spite of the
difference to factor A as factor B changes and (c) shows an extreme interaction where the size of the
response in factor A is reversed as factor B changes.
23
A wide range of factors affect biological organisms and there is need to characterize these effects both
separately and in relation to one another, and thus factorial experiments are of major importance in
agricultural experiments. Factorial experiments should, however be used critically because they are
usually much larger and therefore more complex and costly to conduct that several single-factor
experiments. Also, it is unwise to commit oneself to a larger experiment at the investigations beginning
when several small preliminary experiments may indicate promising approaches.
Example: a plant breeder has collected 30 new varieties from Uganda. He wants to assess the reaction of
these new materials to the local environment. Since the local environment may vary in terms of soil
fertility, moisture levels, etc, the ideal case would have been to test the 30 varieties in a factorial
experiment involving such other variables as fertilizer, moisture and population density. The experiment
however becomes very large as factors other than varieties are added. Even if only three levels of
nitrogen were included, the number of treatments would have increased from 30 to 90. Their size will
mean difficulties in financing, land area, control of soil heterogeneity etc. thus, usually the 30 varieties are
tested under one environment first, then these results are used to discard the obviously poor varieties and
to select a few varieties for a more detailed study. Such a preliminary test, for example may t show that
only five varieties are outstanding enough to warrant further testing. These five varieties could then be put
into a factorial experiment with three levels of nitrogen, resulting in a smaller experiment with 15
treatments. Thus, while a factorial experiment will be more informative than single factor experiments,
practical considerations could limit its use.
The 3*5 factorial treatments (three potato varieties and five nitrogen levels)
Variety Any of the basic designs for single-factor

Nitrogen level experiments can be applied to a factorial
(kg/ha) Asante (V1) p92 (V2) Makonde (V3) experiment. The combination of all the
0 (No) N0 V1 N0 V2 N0 V3 factors involved results in the total number
40 (N1) N1 V1 N1 V2 N1 V3 of treatments and the randomization is
70 (N2) N2 V1 N2 V2 N2V3 done as if the treatments are unrelated.
100 (N3) N3 V1 N3V2 N3 V3
130 (N4) N4 V1 N4 V1 N4 V3
Yield (t/ha) Treatment A two-factor

Nitrogen level Block I Block II Block III Block IV Total Mean experiment in
randomized
Asante
complete block
0 3.852 2.606 3.144 2.984 12.496 3.124 design with four
40 4.788 4.936 4.562 4.608 18.894 4.723 replications
70 4.576 4.454 4.884 3.924 17.838 4.46 Yield of three
100 6.034 5.276 5.906 5.652 22.868 5.717 potato varieties
130 5.874 5.916 5.984 5.518 23.292 5.823 under five levels
p92 of nitrogen:
0 3.852 3.794 4.108 3.444 14.192 3.548
40 4.956 5.128 4.15 4.99 19.224 4.806
70 5.928 5.698 5.81 4.308 21.744 5.436
100 5.664 5.362 6.458 5.474 22.958 5.74
130 5.458 5.546 5.786 5.932 22.722 5.68
Makonde
0 4.192 3.754 3.738 3.428 15.112 3.778
40 5.25 4.582 4.896 4.286 19.014 4.754
70 5.822 4.848 5.678 4.932 21.28 5.52
100 5.888 5.524 6.042 4.756 22.21 5.552
130 5.864 6.264 6.056 5.362 23.546 5.886
Block total 76.992 73.688 77.202 69.508
Grand total 297.39
Grand mean 4.956
24
Source of Degrees of Sum of Mean Observed Tabular Field layout of the 3*5
variation freedom squares square F F factorial experiment in a
Block r-1=3 randomized complete block
Variety v-1=2 design with four replications
Nitrogen n-1=4
Variety*nitrogen (v-1)(n-1)=8
Error (r-1)(vn-1)=42
Total Rvn-1=59
Block I Block III

V3N2 V2N1 V1N4 V1N1 V2N3 V1N1 V3N0 V1N0 V3N1 V1N4
V3No V1N3 V3N4 V1N2 V3N3 V2N2 V1N2 V1N3 V2N4 V3N4
Block II Block IV
Computation steps are: Nitrogen Yield (t/ha)

Nitrogen
Step 1: Compute treatment totals, treatment total
V1 V2 V3
means and the grand total
N0 12.496 14.192 15.112 41.8
Step 2: Let r be the number of replications, V be N1 18.894 19.224 19.014 57.132
the number of variations, and n be the number N2 17.838 21.744 21.28 60.862
of nitrogen levels, construct an outline of the
ANOVA N3 22.868 22.958 22.21 68.036
N4 23.292 22.722 23.546 69.56
Step 3: Enter the 15 treatments totals in the two- Variety total 95.388 100.84 101.162 297.39
way table:
Step 4; Calculate the various sum of squares as follows:

C.F=(grand total)2/total number of observations =(297.390)2/60 = 1474.014
Total SS = x – C.F
2
= [(3.852)2 + (2.606)2 + …… + (5.362)2] – C.F
= 1527.544 – 1474.014 = 53.530
Block SS = (block total)2/vn – C.F = (76.992)2 + …. + (69.508)2 /15 – C.F
= 1476.613 – 1474.014 = 2.599
Treatment SS = (treatment total)2/r – C.F = (12.496)2 + …. + (23.546)2 /15 – C.F
= 1518.592 – 1474.014 = 44.578
Error SS = (total SS – Block SS – treatment SS) = (53.530 – 2.599 – 44.578
= 6.353
Variety SS = (variety total)2/rn – C.F = (95.388)2 + (100.840)2 + (101.162)2 /20 – C.F
= 1457.066 – 1474.014 = 1.052
Nitrogen SS = (nitrogen total)2/rv – C.F = (41.8000)2 + …. + (69.560)2 /12 – C.F
= 1515.248 – 1474.014 = 41.234
variety * nitrogen SS = treatments SS – Variety SS – nitrogen SS = 44.578 – 1.052 – 41.234
= 2.292
Step 5: calculate mean squares by dividing each SS by its d.f.
Step 6: compute observed F-values by dividing the mean square for each source of variation by the error mean
square.
Step 7: look up the corresponding tabulated F-values
Step 8: enter all values obtained in steps 4 –7 in the ANOVA
25
Source of Degrees Sum of Mean Observed Tabular F
variation of squares squares F (5%) (1%)
Block 3 2.599
Variety 2 1.052 0.526 3.48* 3.22 5.15
Nitrogen 4 41.234 10.308 68.26** 2.59 3.8
Variety*nitrogen 8 2.292 0.286 1.89ns 2.17 2.96
Error 42 6.353 0.151
Total 59 53.53
*** - Significant at 1% level; * - significant at 5% level; ns – not significant
SPLIT-PLOT DESIGN
Specifically developed for factorial experiments. Its basic feature is thee use of two plot sizes, one each for the two
variables under study. For example, one for variety and one for nitrogen level. The larger plot is called the main
plot. The number of main plots in a replication corresponds to the levels of the first factor. Each main plot is
subdivided into smaller size plots, called subplots, such that the number of subplots in each main plot corresponds to
the level of the second factor assigned to the subplots.
Using tow plot sizes results in two levels of precision, one for the main plot and one for the subplot. The expected
precision for the main plot is lower than that for subplot. In other words, smaller differences can be detected among
subplots treatments than among main plot treatments.
Consequently, a careful consideration of the relative importance of the factors involved should be made before using
the design. For example, do you want more precision in varietal comparisons or nitrogen level comparisons? The
split-plot design is recommended for use when:
1. The practical limit for plot size is much larger in one factor compared with the other. For example, in an
experiment to evaluate water management and population densities, practical considerations necessitate
using large plots for water managements. Water management levels could then be assigned to the main
plot while population densities are allocated to the subplot.
2. Greater precision is desired in one factor relative to the other. For example, in a study to evaluate the
performance of varieties at several fertilizer levels, if the factor of primary importance is the variety rather
than the fertilizer level, then the fertilizer level should be assigned to the main plot and varieties to the
subplot.
RANDOMIZATION AND LAYOUT

Is a two-stage one. It involves assigning the levels of the first factor to the main plot in accordance with any of the
basic designs learnt earlier. This is followed by random assignments of the levels of the 2 nd factor to the subplots in
each main plot. In other words, there are two separate randomization process: one for the main plots and another for
the subplots. As an example, consider a factorial experiment with six levels of nitrogen as main plots and four potato
varieties as subplots arranged according to a RCBD with three replications. The steps in the randomization are as
follows:
1st divide the experimental area into three blocks of equal size. Each of the three blocks is further subdivided into
six main plots. Starting with the 1st block, allocate at random the six levels of nitrogen fertilizer (N 0, N1, N2, N3, N4,
N5) to the main plots. Repeat the process for the two blocks independently.
Random assignment of nitrogen levels to main plots.
Block I Block II Block III

N4 N3 N1 N0 N5 N2 N1 N0 N5 N2 N4 N3 N0 N1 N4 N5 N3 N2
Note that up to this stage, the randomization scheme is merely one of assigning six treatments with three replications
in randomized complete block manner.
Next, divide each of the 18 main plots into four subplots. Assign the four varieties at random to the four subplots in
each main plot. The layout of this particular experiment is as follows:
26
Block I Block II Block III
N4 N3 N1 N0 N5 N2 N1 N0 N5 N2 N4 N3 N0 N1 N4 N5 N3 N2
V2 V2 V1 V2 V4 V3 V1 V4 V3 V1 V1 V3 V4 V3 V3 V1 V2 V1
Hypothetical layout of a split-plot design for testing 4 potato varieties (subplot) at six nitrogen levels (main plots) in
three replications.
Note that the randomization procedure assures that all the subplots composing a main plot are adjoining each other.
Thus, our example, all four varieties in each vertical strip received the same fertilizer level./ this strip, then, is
considered as only one plot (main plot) for fertilizer treatment, but is composed of four separate plots (subplots), for
varieties.
Analysis of variance:
As an example we look at data on potato yield from an experiment involving six levels of nitrogen and four
varieties. A split-plot design was used with the six levels of nitrogen as main plots and the four potato varieties as
subplots. The main plot treatments were arranged according to the randomized complete block arrangement with
three replications. Thus, the layout would be similar to that above.
Let n represent the levels of the main plot treatment (nitrogen) and v represent the subplot treatment (variety). The
steps involved in constructing the ANOVA of a split-plot design are:
Step 1:
Compute
C.F = (grand total)2 /total no. of observations
= (394481)2/72
= 2161323047
Total SS = x2 – C.F

= [(4430)2 + …+ (2014)2 – 2161323047
= 204747916
Potato yield data for four potato varieties grown under six levels of nitrogen in 3 replications
27
Potato yield (kg/ha) Step 2: construct an outline of the ANOVA
Variety Rep.I Rep.II Rep.III Source of Degrees of Sum of Mean Observed Tabular F
variation freedom squares square F (5%) (1%)
N1(0kg N/ha)
Block r-1=2
Asante (V1) 4430 4478 3850
Nitrogen n-1=5
Tigoni 1 (V2) 3944 5314 3660
Error (r-1)(n-1)=10
Tigoni 5 (V3) 3464 2944 3142 Variety v-1=3
Peru 4126 4482 4836 Nitrogen *variety (n-1)(v-1)=15
N2(60kg N/ha) Total Rnv-1=36
Asante 5418 5166 6432 Step3: the computation of sum of squares will be separated into
Tigoni 1 6502 5858 5586 two parts; main plot and subplot.
Tigoni 5 4768 6004 5556 (a) Main plot analysis. 1st construct the two-way table of main plot
Peru 5192 4604 4652 totals (block*nitrogen take of totals)
N3(90kg N/ha) The block * nitrogen table of yield totals computed from the potato
Asante 6076 6420 6704 yield data
Tigoni 1 6008 6127 6542 Nitrogen Yield total (kg/ha)
Block I Block II Block II N totals
Tigoni 5 6244 5724 6014
N1 15964 17218 15488 48670 (N1)
Peru 4546 5744 4146
N2 21880 21632 22226 35738 (N2)
N4(120kg N/ha)
N3 22874 24015 23506 70395 (N3)
Asante 6462 7056 6680
N4 22167 24954 23252 70373 (N4)
Tigoni 1 7139 6982 6564
N5 23466 23064 23214 69744 (N5)
Tigoni 5 5792 5880 6370 N6 22522 24721 22318 69561 (N6)
Peru 2774 5036 3638 Block total 128873 135604 130004
N5(150kg N/ha) Grand total 394481
Asante 7290 7848 7552 Then compute the following SSs;
Tigoni 1 7682 6594 6576 Block SS = (B2)/nv – C.F
Tigoni 5 7080 6662 6320 = (128873)2 +… + (130004)2/(6)(4)–2161323047
Peru 1414 1960 2722 = 1082577
N6(180kg N/ha) Nitrogen SS = (N)2/rv – C.F
Asante 8452 8832 8818 = (48670)2 + … + (69561)2 /(3)(4) – 2161323047
Tigoni 1 6228 7387 6006 = 30429200
Tigoni 5 5594 7122 5480 Error(a) SS = (NB)2/v – C.F – block SS – nitrogen SS
Peru 2248 1380 2014 = (15964)2 + …. + (22318)2 /(4) – 2161323047 –
1082577 - 30429200
= 1419678
Note that this analysis is exactly the same as that of a RCBD with 3
replications and 6 treatments.
a. Subplot analysis. Construct the two-way table of treatments totals (main plot * subplot)
Nitrogen Yield total (kg/ha)
V1 V2 V3 V4 N totals
N1 12758 12918 9550 13444 48670 (N1)
N2 17016 17946 16328 14448 65738 (N2)
N3 19200 18777 17982 14436 70395 (N3)
N4 20196 20685 18042 11448 70373 (N4)
N5 22690 20852 20062 6140 69744 (N5)
N6 26102 19621 18196 5642 69561 (N6)
Block total 117964 110799 100160 65558
Grand total 394481
Then, compute the following SSs:
Variety SS = (v2)/rn– C.F
= (117964)2 +… + (65558)2/(6)(3)–2161323047
= 89888101
Variety*Nitrogen SS = (vN)2/r – C.F – variety SS – nitrogen SS
= (12758)2 + … + (56421)2 /(3) – 2161323047 – 89888101 - 30429200
28
= 69343487
Error(b) SS = Total SS – sum of all other SS
=204747916 - (1082577 + 30429200 + 1419678 + 89888101 +69343487)
= 12584873
step 4: compute mean squares for each source of variation by dividing the sum of squares by its corresponding
degrees of freedom as follows:
Nitrogen MS = 30429200/5 = 6085840
Error(a) MS = 1419678/10 = 141968
Variety MS = 89888101/3 = 29962700
Nitrogen*Variety MS = 69343487/15 = 4622899
Error(b) MS = 12584873/36 = 349580
Step 5: compute the observed F-values. The block and main plot(in this instance, nitrogen effects are tested usung
error(a) while the subplot treatment (variety) and the interaction effects are tested with error(b). thus
F for nitrogen = 6085840/141968 = 42.87
F for variety = 29962700/349580 = 85.71
F for interaction = 4622899/349580 = 13.22
Step 6: compute the coefficients of variations. Since the split-plot analysis has two error terms, there are also two
coefficients of variation to be computed, one for the main plot analysis, cv(a) and another for the subplot analysis
cv(b).
cv(a) = (error (a) MS/grand mean) * 100 = (141968/5479)*100 = 6.9%

cv(b) = (error (b) MS/grand mean) * 100 =(349580/5479) * 100 = 10.8%
the cv(a) indicates the degree of precision attached to the measurement of the main plot treatment effects and the
cv(b) the precision of the subplot treatment and interaction effects.
Usually we would expect cv(b)to be smaller that cv(a) since as indicated earlier, the factor assigned to the main plot
is measured at relatively lower precision than those assigned to the subplot. This trend does not always hold, as
shown by the present example, and one should consult a competent statistician if such results frequently occur in
your experiments.
Step 7: Enter all values obtained from steps 1 to 6 in the analysis of variance table.
Source of Variation Degrees of Sum of Mean Observed Tabular F
freedom squares square F 5% 1%
Block 2 1082577 541228
Nitrogen 5 30429200 6085840 42.87** 3.33 5.64
Error (a) 10 1419678 141968
Variety 3 89888101 29962700 85.71** 2.86 4.38
Nitrogen *Variety 15 69343487 4622899 13.22** 1.96 2.58
Error (b) 36 12584873 349580
Total 71 204747916
cv(a)=6.9%; cv(b)=10.8%, **=significant at the 1% level
Compare the observed F-values with their corresponding tabular values and indicate its significance by appropriate
asterisk. In this example, all the three effects (N, V and N*V) are highly significant. When the interaction is
significant, the interpretation of the results becomes complicated, and mean comparison for factorial treatments
should be done.
CHI – SQUARE TEST.

Most agricultural research data can be classified as either measurement data or attribute data. Measurement data
refer to data recorded on some numerical scale, but attribute data classify experimental units into distinct groups
based on some qualitative characters. Examples of measurement data are grain yield, plant height, number of tillers
etc. Attribute data is usually recorded in ‘words’ instead of numbers e.g. color classification, farmer’s tenure status,
marital status, sex etc.
Examples of agricultural experiments where attribute data rather than measurement data answer more adequately
certain experimental objectives are:
29
1. A plant breeder is studying across between a maize inbred line with white endosperm and another with
yellow endosperm. He would like to know whether the ratio of endosperm colour in the F 2 population
follows a 3:1 ratio. From the cross he obtains F2 kernels and classifies according to the two colour
categories. Suppose from 400 F2 kernels examined, he finds 325 that are yellow and 75 that are white. He
then asks; does the observed ratio of 325:75 deviate significantly from the hypothesized ratio of 3:1?
2. A certain insect, green grasshopper is suspected to have a different preference toward feeding on an all
ready diseased plant or undiseased plant. The researcher, therefore inoculates a few plants and then subjects
these injected plants together with healthy plants of the same variety grown under uniform management, to
a common destiny of insect population. He then determines whether the proportions of insects found on
diseased plant is the same as that on healthy plants i.e. whether the ratio of insects found on diseased and
healthy plants is 1:1.
3. An agricultural economist studying factors affecting the adoption of newly introduced high-yielding rice
varieties wishes to know whether adoption is affected by the tenure status of farmers. Since there are 3
distinct tenure statuses, namely; owner operator, leaseholder and tenant farmer, he then proceeds to
determine whether the ratio of adopters to non adopters remain the same over the three categories of
farmers based on tenure status.
USES OF X2 TESTS
Three common uses are:
1. Test for a fixed ratio hypothesis: - many agricultural experiments are done to verify empirically some
biological phenomena that are predicted to occur given that some assumptions are satisfied. This type of
experiment is common in inheritance studies as typified by example 1 in the previous section e.g. a ratio
of 3:1 is expected to occur in the F2 of a cross between yellow and white endospermed maize inbred.
Whether expectation is substantiated by the actual data can be answered by a X 2- test. Note that this type
of X2 – test can be used regardless of the number of classes provided that an expected ratio is specified
for all classes before experimentation.
2. Test independence in a contingency Table:- when attribute data are classified according to two criteria,
each with several classes, we have an R*C contingency table, where R is the number of classes in the 1st
criterion and C, the number of classes in the 2 nd .
Example 3 in the previous section illustrates a 2*3 contingency table where the two criteria are adoption status with
two classes and tenure status with three classes. The question relevant to this type of problem is whether the ratio
among classes in one criterion is affected by the classes of the other criterion. For the adoption example, the relevant
question is whether the proportion of adoption is affected by the tenure status of the farmers. The null hypothesis in
this instance is that the two criteria are independent of each other and the X 2 – test can be for such a null hypothesis
is known as the X2 – test of independence.
3.Test of goodness of fit: - although the X2 – test is primarily used for attribute data, it can also be helpful
in testing if a set of measurement data follows a hypothesized probability distribution. In analysis of
variance it is assumed that the data follow the normal distribution. If certain data are suspected to violate
this assumption the X2 – test can be used to determine whether the normal distribution fits the data
sufficiently well. The goodness of fit of any set of data to any form of probability distribution can be tested
by a X2 – test.
COMPUTATIONAL PROCEDURES
The procedure for computing a X2 – test is very similar for all the three uses mentioned, and the steps involved are:
Step 1: determine the sample size or the number of experimental (or sampling) units that have been classified into
various attributes. In example 1, since 400 kernels were examined for colour, the sample size is 400.
Step 2: determine the number of criteria upon which each sampling unit is being classified. This step specifies
whether the data belong to a one-way classification, two-way classification, or other multi-way classification. For
example, in example1 colour is the only criterion being considered so that we have a one-way classification with
two cells, white or yellow kernels. For example 3, on the other hand, the data belong to a two-way classification
problem since each farmer is classified according to the both adoption status and tenure status. Furthermore, in this
example the number of cells, or the number of distinct groups to which each unit belongs, is 2*3 or 6, namely;
owner adopter, owner non-adopter, tenant adopter, tenant non-adopter, lessee adopter, and lessee non-adopter. The
total number of cells in a contingency table is equal to the product of the number of classes of each criterion.
Step 3: determine the ‘expected value’ for each cell based on the hypothesis set up by the researcher. For example 1,
with sample size of 400 and based on the hypothesis that the ratio between yellow and white kernel is 3:1, the
expected value of the yellow kernel cell is ¾*400 = 300 and the expected value of the white kernel cell is ¼*400 =
100.
Step 4: the actual number of units classified into each cell is designated as the ‘observed value’ of the particular cell.
For example 1, the observed value of the yellow kernel cell is 325 and 75 for the white kernel cell.
30
Step 5: compute the X2 – test as:
X2 = pi=1 (observed  – expected )2 /expected 

Where p is the total number of cells; observed  is the observed value and expected  is the expected value of the th
cell. For example 1, the X2 – test is computed as:
X2 = (325 – 300)2 /300 + (75 – 100)2 /100
= 8.33
Step 6: compare the observed X2 – value computed in step 5 with the tabular X2 – value obtained from the tables.
The degreees of freedom (d.f) to be used for choosing proper X2 – values are determined as follows:
a) For one-way classification problem, the d.f is p – 1 where p is the total number of cells.
b) Fro two-way classification with an r*c contingency table the d.f is (r – 1)(c – 1), where r is the number of
rows and c is the number of columns of the two-way table.
c) For higher than two-way classifications, say with k classification criteria, the general rule is that the d. f =
(P1 –1) (P2 –1) (P3 –1)….. (Pk –1), where Pi is the number of classes in the Ith classification criterion.
If the observed X2 – value is greater than the corresponding tabular value, the hypothesis is rejected. For example 1
the tabular value with (p – 1) = (z – 1) = , degree of freedom is 3.84 for 5% level of significance and 6.63 for 1%
level of significance. Since the observed X2 – value of 8.33 is greater than the tabular values at 5% level and 1%
level, the test is highly significant. Hence, the hypothesis is rejected. That is, the data did not support the
hypothesized ratio of 3:1 between yellow and white kernel in the F2 population.
Distribution of χ2 for 2, 4, and 6 degrees of freedom

Obviously χ2 cannot be negative since it involves a sum of square numbers.
TEST FOR INDEPENDENCE IN A CONTINGENCY TABLE

Plants breeders of KARI investigated the resistance to citrus aphid and resistance to greeing disease in oranges. An
aphid resistant line of KA98 was crossed to a greening disease resistant line of KA01 from which a total of 374F 3
progenies were derived and evaluated. Data below: Number of F 3 progenies from a cross between an aphid resistant
line and a greening disease resistant line classified into various degrees of resistance:
F3 progenies (no) classified
Aphid resistant reaction according to greening disease Total
Resistant Segregating Susceptible
Resistant 24 49 23 96
Segregating 53 74 45 172
Susceptible 20 61 25 106
Total 97 184 93 374
For this experiment the questions that need to be answered and the tests to answer them are as follows:
Is the reaction to aphids independent of that of greening disease? This is equivalent to the question: Does the
proportion of F3 lines classified into three classes og greening disease reaction (i.e. resistance, segregating and
susceptible) remain the same for various classes of aphid reaction and vice versa? Note that the question is asked
without making any assumption on the tru proportion of each reaction. To answer this question, a X 2 – test for
independence is needed. The steps involved are:
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Step 1: compute the expected value for each of the nine (3*3) cells;
Eii= ni.n.i /n..
Where Eij is the expected value of the I,jth cell, n1. and n.1 represent the totals of the 1th row and jth column,
repectively, and n.. is the grand total. For example, the expected value for the first cell is computed as:
E11=n1.n.1/n.. = (96)(97)/374 = 24.9
Following are the results of all the nine cells.
F3 progenies (no) classified
Aphid resistant reaction according to greening disease
Resistant Segregating Susceptible
Resistant 24.9 47.2 23.9
Segregating 44.6 84.6 42.8
Susceptible 27.5 52.1 26.4
Step 2: compute the X2 – test as follows:
X2 = (observed – expected)2/expected
= (24 – 24.9)2/24.9 + (49 – 47.2)2 /47.2+ …..+(25-26.4)2/26.4
= 6.80
Step 3: Compare the observed x2-value of 6.80 with the tabular x2-value with (r - 1)(c –1)=4 degrees of freedom (i.e.
X2=9.49 at 5%level of significance). Since the observed X 2-value is smaller than the corresponding tabular value at
5% level of significance, the hypothesis of independence cannot be rejected, i.e. there is insufficient evidence to
conclude that the greening disease reaction and the aphid resistance reaction are dependent on one.
COMPARISONS AMONG TREATMENT MEANS

The significant F-test in the ANOVA indicates the presence of one or more real differences the treatments tested.
But it does not locate the specific difference or differences that may account for the significance. Thus the ANOVA
should be considered only as the first step in evaluating differences among treatments. The step that follows is to
locate the specific treatment differences of interest to the researcher and test whether these differences are
significant.
Locating and testing specific treatment comparison:
Specific comparison among treatment means can be arbitrarily classified into three types:
1. Comparisons between pairs of treatment
2. Comparisons between two groups of treatments
3. Trend comparison
The first is the simplest since each comparison involves only two treatments. Because of its simplicity, this
comparison is the most common used.
The 2nd comparison involves classifying the treatments into meaningful groups. Each group may consist of one or
more treatments nad comparison is made between the aggregate means. One example of this is a comparison
between the mean of all fertilized plots and the mean of unfertilized plots; another is the comparison between the
mean of several native varieties and the mean of several of several improved or newly introduced varieties.
While the 1st two comparisons can be applied to any set of treatments, the third or trend comparison is limited to
treatments that are quantitative, for example, rates of fertilizer and distances of planting. Although the trend
comparison is slightly more complicated, it can also generate more information than can be obtained from the 1 st
two comparisons.
Comparison Between Pairs of Means: two ways of comparing pairs of treatment means are
a) Preplanned comparisons: - based on pairs of treatments that have been specifically planned and pinpointed
before the start of the experiment e.g. comparing each of the several treatments to a control, as in
comparing the unfertilized treatment to each of the fertilized treatments.
b) Unplanned comparisons: - no specific comparison is chosen in advance. Instead all possible comparisons
are evaluated to see which difference may appear real.
Two of the more commonly used procedures are the least significant difference test and Duncan’s Multiple Range
Test.
Least Significant Difference (LSD) – simple, and the most commonly used procedure for evaluating the significance
of difference between pairs of treatment means. All that is done to locate any pairs of means whose difference
between pairs of treatment exceeds the LSD value and declare the means significantly different from each other.
Strictly speaking, however, this procedure is valid when the experiment involves only two treatments, because as
32
more treatments are involved and when all possible pairs of treatments means (unplanned comparisons) are to be
tested the LSD becomes less and less precise.
Note that the number of all possible pairs increases very rapidly as the number of treatments increases i.e. 10
possible pairs for 5 treatments, 45 for 10 treatments and 105 for 15 treatments. Furthermore it can be shown that the
probability of at least one comparison, the largest versus the smallest, exceeding the LSD value at 5% level of
significance when, in fact the difference is not real, is 29% for 5 treatments, 63% for 10 treatments and 83% for 15
treatments. This implies that when the experimenter thinks he testing at the 5% level of significance, he is actually
testing at 29% level of significance for 5 treatments, 63% for 10 treatments and so on. Thus, the LSD test can be and
is often misused. Hence, special care must be exercised in using this test. The following rules have been found
useful in the effort to use the LSD test effectively:
a) Use the LSD test only when the F-test in the ANOVA is significant
b) Do not use the LSD test for comparisons among all possible pairs of means when experiment involves
more than 5 treatments
c) Use the LSD test for preplanned comparisons regardless of the number of treatments involved.
For instance, in comparing every treatment to a control, the LSD test can be used even if there are more than 5
treatments.
LSD values for experiments with equal replications – computational procedures;
When every treatment is replicated r times, the formula for calculating the LSD value at a certain level of
significant, say , is LSD = t2S2/r, where t is the tabular value of t at the  level of significance, and with the
error degrees of freedom, S2 is the error mean square from the analysis of variance and r is the number of
replications.
To judge whether a difference between two treatment means is statistically significant, compare
the observed difference with the computed LSD value. If the observed difference is larger than
the LSD value, the two treatment means are significantly different at  level of significance. If
the difference between any pair of means is smaller than the LSD value, the two treatments are
not significantly different.
Example.
As an illustration data from the insecticide test presented in the CRD ANOVA previously will be used. In this
instance, the LSD test will be applied for the comparisons between the control and each of the six insecticide
treatments. This is a planned comparison; hence the used of LSD test is appropriate regardless of the number of
treatments involved.
Information needed for the computation of LSD value is taken from the ANOVA table for this data. Using the
formula LSD = t2S2/r ; LSD values at 5% and 1% levels of significance are computed as
LSD = t2S2/r
LSD = t0.52S2/r
= 2.0802(94773/5 = 453 kg/ha
and
LSD = t0.12S2/r
= 2.831(94773/4 = 616 kg/ha
The observed difference between each of the treatment means from the control mean is:
Comparison between yield of each of the 5 insecticide treatments and control using LSD test (data CRD ANOVA):
Treatment Difference To judge which differences are statistically significant,
mean from Control each difference is compared with the computed LSD
Treatment (kg/ha) (kg/ha) values. If it exceeds the LSD value only at the 5% level,
Malathion (1kg) 2127 811** one asterisk is used. Otherwise use an ns to indicate ther
Malathion (2kg) 2678 1362** difference is not significant. Of the six insecticides, only
Thiodan 2552 1236 ‘Karate’ was not significantly different from the control.
Diazinon-A 2128 812** Others are significantly superior either at 5% level or at
Diazinon-B 1796 480* 1% level.
Karate 1681 365ns
Control 1316 ---
** = Significant at 1% level, * = significant at 5% level,
ns = not significant
DUNCAN’S MULTIPLE RANGE TEST (DMRT)
33
In contrast to the LSD test in which only values is computed for all comparisons, the Duncan’s multiple range test
prescribes a set of significant differences of increasing sizes depending upon the distance between the rankings of
the two means to be compared. While additional computations are required for the DMRT, the test overcomes the
major defects of the LSD test in that DMRT can be used to test differences among all possible pairs of treatments
regardless of the number of treatments involved and still maintain the prescribed level of significance. The steps in
computing the DMRT are:
Step 1: Arrange the treatment means in decreasing or increasing order
Step 2: Calculate standard error of a treatment mean, as follows S x = S2/r, where S2 is the error mean square and r
is the number of replications
Step 3: Calculate the ‘shortest significant ranges’ for various ranges of means as follows:
Rp = rpSx , where rp)=2, 3 ..t) are the values of ‘significant studentized ranges’ obtained from the tables based on the
error degrees of freedom and t is the number of treatments.
Step 4: group the treatment means according to the statistical significance. For this, the following method may be
used:
a) From the largest mean, subtract the ‘shortest significant range’ of the largest p. declare all means less than
this value significantly different from the largest mean. For the remaining means not declared significantly
different, compare the range i.e. difference between the largest and the smallest, with appropriate Rp. If the
range is smaller than its corresponding Rp, all remaining means are not significantly different.
b) From the second largest mean, subtract the second largest Rp. declare all means less than this value
significantly different from the second largest mean. Then, compared the range of the remaining means
with the appropriate Rp.
c) Contnue the process with the third largest mean, then the fourth, and so on until all means have been
properly compared.
The steps in the computation of Duncan’s multiple range test will be illustrated using data in the CRD ANOVA.
Step 1: Arrange the means in decreasing order as follows:
Mean yield Step 4: the following steps should be followed in
Treatment (kg/ha) Rank grouping the means according to statistical significance.
T2 Malathion (1kg) 2678 1 1. From the largest treatment mean of 2678 kg/ha,
T3 Malathion (2kg) 2552 2 subtract the largest Rp (i.e. R7 = 513) and
T4 Thiodan 2128 3 obtain the difference of 2165 kg/ha. Declare all
T1 Diazinon-A 2127 4 treatment means less than 2165 as significantly
T5 Diazinon-B 1796 5 different from the largest mean. From the array
T6 Karate 1681 6 of means shown in step 1, all treatment except
T7 Control 1316 7
T3 are significantly different from T 2. Since
there are two remaining means, namely T 2 and
Step 2: Calculate standard error of a treatment mean as: -
T3, either difference of 2678 – 25552 = 126
Sx = S2/r = 94773/4 = 153.93 kg/ha
kg/ha is compared to R2 = 453. Since the
difference is smaller than R2, T2 and T3 are not
Step 3; from the tables with error d.f of 21 obtain the
significantly different.
values of ‘significant studentized ranges’ for the 5%
Draw a vertical line connecting the means of T 2 and
level of significance as follows:
T3 as follows:
P rp(0.05) Treatment Mean yield
2 2.94 (by rank)
3 3.09 T2 2678
4 3.18 T3 2552
5 3.25 T4 2128
6 3.3 T1 2127
7 3.33 T5 1796
Then compute the ‘shortest significant ranges’ using
formula Rp = rpSx T6 1681
T7 1316
1. The vertical line is a convenient and widely
accepted way of marking differences among
treatment means. Note that all means connected
by the same line have been judged not
significantly different from each other.
2. 2. From the 2nd largest mean (T3) of 2552
34
P Rp=rpSx kg/ha, subtract the second largest Rp(i.e. R6)
i.e. 2552 –508 = 2044. Declare all the means
2 2.94(153.93) = 453 less than 2044 kg/ha significantly different
3 3.09(153.93) = 476 from the mean of T3. Here, the means of T5, T6
4 3.18(153.93) = 490 and T7, are less than 2044 kg/ha and hence are
significantly different from T 3.
5 3.25(153.93) = 500
6 3.30(153.93) = 508
7 3.33(153.93) = 513
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The range of the remaining means that were not
declared significantly different is T 3 – T1 = 2552 – The only difference that is not significant is between
2127 = 425 which is less than R3 = 476. Hence, all T6 and T7. Hence a common vertical line connects
remaining means namely T3, T4 and T1 are not these two means as follows:
significantly different. Draw a vertical line Treatment Mean yield
connecting the means T3, T4 and T1 as follows.
(by rank)
Treatment Mean yield
T2 2678
(by rank)
T3 2552
T2 2678
T4 2128
T3 2552
T1 2127
T4 2128
T5 1796
T1 2127
T6 1681
T5 1796
T7 1316
T6 1681
T7 1316 In the presentation of the results, the use of vertical lines
3. From the third largest mean (T 4) of 2128 kg/ha, significant differences is appropriate only when the
subtract the third largest Rp of 500kg/ha to treatment means are arranged according t rank.
obtain the difference of 1628 kg/ha. Declare all Otherwise, letters should be substituted for the lines: -
mean less than 1628 kg/ha significantly
different from T4. Here, T7 with a mean of 1316 Treatment Statistical
Treatment1
kg/ha is significantly different from T 4. The 2
mean (kg/ha) Significance 2
range of the remaining four means (T 4, T1, T5
and T6) is 2128 –1681 = 447kg/ha which is less T1 2127 bc
than R4 = 490. Thus, there is significant
T2 2678 a
different among the remaining means. Draw a
vertical line connecting the four means as T3 2552 ab
shown. T4 2128 bc
Treatment Mean yield
T5 1796 c
(by rank)
T2 2678 T6 1681 cd
T3 2552 T7 1316 d
1
T4 2128 Average of four replicates
2
any two means having a common letter are not
T1 2127
significantly different at 5% level.
T5 1796 -the letter designation means that any two means having
T6 1681 at least one common letter are not significantly different.
T7 1316 Line designation is transferred to letter designation
simply by replacing each line with a letter. In this
4. Here the process can be continued with the
instance there is a total of four lines, hence four letters
fourth largest mean and so on. However, since
(a, b, c, and d) are used.
T7 is at this stage, the only mean outside the the
groupings all ready made, it is simpler just to
compare T7 with the rest of the means (namely
T1, T5 and T6) using the appropriate Rp. The
comparisons are
T1 – T7 = 2127 – 1316 = 811 >R4 = 490
T5 – T7 = 1796 – 1316 = 480 >R3 = 476
T6– T7 = 1681 – 1316 = 365 < R2 = 453
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MECHANICAL ERRORS
Result from faulty execution of experimental procedures, many of which are human mistakes that could
make a substantial difference when committed during initial periods of the experiments. Statistical tools
are less effective in coping with mechanical errors, unlike other sources of error, such as soil
heterogeneity and competition effect.
Furrowing - for row crops any errors in row spacing are reflected as error in plot measure. For a sorghum
varietal test sown on rows 0.75m apart, a plot size of 4 rows 5m long would measure 3 * 5m. If the
spacing between rows in one plot were incorrectly set at 0.80m, instead of 0.75m intended. Then a plot
size of 3.2 * 5m would result. And the plot performance would definitely be affected such a difference in
the area size planted.
Faulty furrowing in directly seeded row crops usually causes errors in row spacing. Despite precautions
being taken, at times anomalies in row spacing are not discovered early enough for re-furrowing. When
this happens, two alternatives are subjected:
1. Mathematically correct the plot yield based on the discrepancy in area between the normal and
anomalous plot
2. Harvest only from portions of the plot where row spacing is correct.
SELECTION OF SEEDLINGS
For transplanted crops, seedlings are first raised in seedbeds before being transplanted to the
experimental area. To assure sufficient plants, more seedlings are usually grown than are actually
required. This allow for some type of seed selection. The usual practice in transplanting is to use first the
best looking seedlings, leaving the poor seedlings as leftovers.
The basis for selection should be uniformity and not maximum vigour.
Thinning – for most directly seeded crops high seeding rate is used to ensure that enough seeds will
germinate. Several days after germination, however, the number of plants per plot is reduced to a
constant number by pulling out (thinning) the excess plants. Some of the more common problems in the
thinning process that could increase experimental error if not properly handled are:
1. Selecting plants to be retained – The decision is complex for crops drilled in the rows because apart
from trying to retain the best looking seedlings, plants have to be kept at the prescribed spacing. In
sorghum, for example, the prescribed distance between plants in the row is after thinning is 10cm. At
this spacing, it is not easy to determine visually which plants to retain and which to remove. Usually a
worker is provided with a stick about the length of a plot row that is properly marked every 10cm. The
worker then sets this stick beside every row to be thinned and only plants situated closest to the
10cm marks are retained.
2. Presence of vacant spaces in the row. It is not uncommon in row-sown crops to find spots in a row
with no plants. In such instances, the thinning process may be manipulated so that areas adjacent to
the vacant space are left with more plants. For example, if there is a vacant space of about 30cm in a
row of sorghum that is to be thinned to one plant per 10cm, the researcher can retain one plant more
than required in the normal thinning at each end of the gap. The resulting locations of the plants in a
1m row will be as indicated.
37
3. Presence of ridges with less than intended for. For directly seeded crops despite all precautions to
assure enough seedlings per ridge, it is not uncommon to have some ridges with fewer plants than
needed. A researcher can do things about this:
a) He can ignore it at the seedling stage and then adjust to the same number of plants at
harvest.
b) He can attempt to correct the problem at the seedling stage by thinning. The procedure is
to retain more plants per ridge in the adjacent ridges so that the desired plant population
is obtained. For, example, two plants per ridge is normally retained in maize performance
test. If a ridge has only one plant, 3 plants could be retained in the next ridge.
4. The correct stage of thinning. Thinning should be done before any tillers are formed so that counting
of primary seed stales is not confused with tillers. Another consideration is the incidence of serious
seedling pests and diseases. For experiments not primarily concerned with evaluating pest and
disease reaction. You could allow the seedlings to pass through the most susceptible stage before
completing final thinning. In sorghum shoot fly is a serious seedling pest. Sorghum however develops
resistance to it sown after the seedling stage. During planting when the pest is serious, thinning can
be done more than once. The first thinning reduces the variability in density of seedlings within amd
among plots while still allowing for flexilibity in correcting for seedling damage in the susceptible
period.
Transplanting – for transplanted crops, the number of seedlings per ridge should be uniform. In most
vegetable crops this is easy since seedlings are usually large and easily separated from each other.
Fertilizer application – uniform application of fertilizers, even in large areas, is not much of a problem if
mechanized fertilizer spreaders are available. In Kenya, however, fertilizer application is done normally
and uniformity is difficult to ensure, especially when large areas are concerned. It is almost impossible to
fertilize a hectare uniformly by hand, but is a much simpler task to do so when only, say, 10 to 50 square
metres is involved. Therefore, the experimental area should be subdivided into smaller units before
fertilizer is applied. The field worker carries with him bulled fertilizer in one hand and a volume measure
(empty can of milk, tablespoon etc) in another. He then fills the measuring container, spreads it over
specified unit area, moves to the next unit, fills the same can and empties it over this area, and so on.
COMPETITION EFFECTS
An individual plant performance is greatly affected by the side and proximity of adjacent plants.
Individuals surrounded by large and vigours ones. Further more, plants a greater distance from one
generally produced more that those nearer to each other. The interdependence of adjacent plants
because of their common need for limited solar energy, soil nutrients, moisture, CO 2 etc is commonly
referred to as competition effects.
TYPES OF COMPETITION EFFECTS
For a given experiment, the significance of any competition effect depends primarily on the treatments
being tested and the experimental layout.
Varietal competition- in evaluating the performance of different varieties of given crop, adjacent plots are
necessarily planted to different varieties. Varieties generally differ in their ability to compete, however and
plants in a lot will not be subjected to the same environment depending upon their relative location
adjacent plots. This is known as varietal effects. The plants generally affected by this competition are the
ones near the plot’s perimeter.
Fertilizer competition – is a similar to the varietal competition except that adjacent plots receive different
levels of fertilizer instead of being planted to different varieties.
The competition is a result of two contrasting sources:
Plots with higher fertilizer application will be more vigorous and therefore compete better for solar energy
and CO2.
The fertilizer given to one plot could spread to the root zone of the adjacent plot putting the plot with
higher fertilizer at a disadvantage. These two contrasting effects have their difference constituting the net
competition effect.
Unplanted borders – are areas between plots or around the experimental areas left unplanted to serve as
markers or pathways. Since these areas are generally wider than the area between rows or between
38
plants in a row, plants adjacent to these unplanted borders have relatively more space and are therefore
exposed to les competition pressure compared with those in the plot’s center.
Missing hills – a spot in an experimental plot where a living plant is supposed to have been located but is
absent because of poor germination, pest or disease damage etc is called a missing hill. A hill’s absence
causes the plants surrounding the position where the hill was supposed to have been (i.e. missing hill) to
be exposed to less competition than the other hills. These plants are therefore more favored and usually
perform better than those surrounded by living plants.
ACCURACY AND PRECISION
Accuracy refers to the closeness with which a particular measurement can be made.
Precision refers to the magnitude of the difference between two treatments that an experiment is
capable of detecting.
Accuracy in data collection and compilation – whenever possible, original records should be collected
in a manner to avoid recopying. This avoids possible errors in recopying. If figures must be
transferred, they should always be rechecked immediately. At the time data ids collected it should be
examined for out of line figures and all such entries rechecked to avoid possible errors. There is
enough inherent variation in biological data without allowing more to creep in through human error.
How close should measurement be taken? Do you weigh to the closest hundredths, tenth, or whole
kilogram? Spurious accuracy should be avoided. It is unnecessary to record figures not warranted by
precision of an experiment. A general guide for the number of significant figures to retain is as
follows:
If the coefficient of variability is between 0.4 and 4%, record original data to 4 significant figures; if the
cv is between 4 and 40% carry 3 figures and if the cv is greater than 40%, only record 2 figures.
A more precise rule is that rounding interval should not exceed one-fourth of the standard deviation
per plot. If S is estimated to be 12.50, then ¼(12.5) = 3.1 therefore, record the data to the nearest
whole unit. If S = 2.5, ¼(2.5) = 0.6, record to closest 0.1. If S = 0.20, ¼(0.20) = 0.05, record to the
closest 0.01.
The instrument used for weighing or measuring need be no more accurate than required by the
precision of the experiment. Top make a series of weighing which will be rounded to the nearest
kilogram, the scales to be used can be in whole kilogram units rather than division of a kilogram.
In the analysis of variance, it is best to carry the full number of figures obtained from the uncorrected
total sum of squares. If original data contains one decimal the sum of squares will contain 2 decimal
places.
In the final presentation o results superfluous digits should be avoided. Round treatment means to
one-tenth of the estimated standard error of a mean. If S x = 2.56, 0.1(2.56) = 0.2, therefore, round
treatments to the closest 0.1. Thus, a mean of 15.12 is given as 15.1.
The greater the variability among plots treated alike, the greater will be the error associated with the
difference between two means and the less precise the experiment will be in detecting differences
due to treatments. The standard error of the difference between two means decreases as S
decreases and n increases Sd = t2S2/n (where n is the number of replications). Thus, methods to
increase the precision of an experiment are designed to lower the unaccounted variability per plot or
to increase the effective number of replications.
Precision may be improved by:
1. Increased replication
2. Careful selection of treatments
39
3. Refinement of technique
4. Selection of experimental material
5. Selection of experimental unit
6. Taking additional measurements
7. Planned grouping of experimental units
Increased replication – the precision of an experiment can be increased by additional replications

but the degree of improvement falls off rapidly as the number of replications increases. For
example, compared to an experiment with four replications, to double the degree of precision with
which two means can be separated requires 16 replications. This follows from the effect of the
number of replications (n) or the difference required to separate two means at a given level of
significance, LSD = t2S2/n.
In general, in field and vegetable crop research, from 4 to 8 replications are required for
reasonable precision. In planning an experiment, you should be reasonably sure that you would
be able to detect a true difference of the magnitude in which you are interested.
Selection of treatments - careful selection of treatments is important in achieving the experimenter’s

objectives and increasing the precision of the experiment. Fro example, in studying the effect of a
herbicide, fungicide, fertilizer or insecticide, it is more useful to determine how the experimental units
respond to increasing doses of your treatment, than deciding whether or not two succeeding doses are
significantly different.
Refinement of technique – faulty techniques may increase experimental error and bias treatment effects.
Good technique should aim to:
a) Uniformly apply treatments
b) Devise suitable and unbiased measurement of treatment effects
c) Prevent gross errors
d) Control external influences so that all treatments are comparably affected.
Selection of experimental material - for certain kind of studies, careful selected, uniform material is
desirable. In selecting experimental material keep in mind the population about which you wish to make
inference. It is important to use the kinds of experimental materials that will be used in actual production.
Selection of the experimental unit- size and shape of the field plot affects precision. Generally, variability
decreases with an increase in plot size, but once a certain size has been reached, the increase in
precision falls off rapidly with larger sizes. For determining yield there is no increase in precision by using
plots larger than 0.1 acre. For most crops, harvested areas of 0.01 to 0.02 acres result in good precision.
Rectangular plots are most efficient in overcoming soil heterogeneity when their long axes are in the
direction of greatest soil variation.
Taking additional measurements – a technique known as the analysis of covariance can sometimes be
used to remove an important source of variation among the experimental units. Initial rates growth of
plants in fertilizer experiment might be used to remove this effect on the rate of gain over a fertilization
period.
Planned grouping of experimental units – involves the use of the principles of experimental design we
have designated local control. By certain restrictions on the randomization of treatments to experimental
units it is possible to remove certain sources of variation such as changes in soil fertility across an
experimental area. The grouping of experimental units in different ways gives rise to various experimental
designs.
40
SAMPLING
-plot size in field experiments is chosen to satisfy a prescribed degree of precision for the character of
primary interest. The character of primary interest is usually economic yield (ie grain yield for grain
crops, cane yield for sugarcane, etc) whose characters are also the most difficult to measure, and therefore
the plot size is often larger than needed to measure other characters. Unnecessary expense and time can be
saved if only a fraction of the plot (ie a sample) is used to measure supplementary characters. For
example, in evaluating plant height, measurements may be made from only three of 100 plants in a plot;
for number of leaves only 20 of 500 leaves; etc. Sometimes the choice of plot size and shape may be
governed by the cultural and management practices, which could also result in a much larger plot size
than required even for yield. In such instances, even yield may be determined by sampling.
-an appropriate sample estimates as closely as possible the value that would be obtained if all plants in the
plot were measured. The difference between the sample value and the plot value constitutes the sampling
error. A good sampling procedure must give a small sampling error.
Basic Concepts In Sampling:
Population and Sample- for sampling in replicated trials, each experimental plot is a population. The
population value is the plot value, which could be estimated from a few sample plants taken from each
plot.
Sampling Unit- is the unit in which actual measurement of a character is made. For replicated trials where
the plot is the population, the sampling unit must be smaller than the plot. Some commonly used units
are: a leaf, a plant, a group of plants, and a specified area. A good sampling unit must be (1) easy to
identify (2) easy to measure (3) fairly uniform.
-the best sampling unit is one that will give the highest precision at a reasonable cost. Precision is usually
measured by the variance of the estimate of the character of interest and the cost based on time spent in
measuring each sampling unit. The smaller the variance, the more precise is the estimate. The faster the
measurement process, the lower the cost.
Sample size- refers to the number of sampling units to be measured in each population. For example, in
replicated trials, sample size specifies the number of plants per plot to be measured for height, the number
of leaves to be measured for leaf area, and the number of hills on which tillers will be counted. The choice
of sample size depends on the size of the variability of the character to be measured and the level of
precision required of the estimate.
The treatment mean is the estimate of primary interest in replicated trials. The variability of a character
therefore, is described as the variance among the plots of the same treatment (Vp) and the variance among
sampling units in the same plots (Vs)
-the level of precision is decided by the researcher based on previous experience and his experimental
objective. The researcher prescribes the margin of error in the estimate of a treatment mean that he is
willing to tolerate (eg that the estimate should not deviate from the true value by more than a specified
percent). Having estimated the variance and specified the margin of error, the required sample size for a
random sampling scheme can be estimated as:
n= (Zα2/rd2)(Vs/Ỹ2 )
1-(Zα2/rd2)(Vp/Ỹ2 ) where n is the sample size, r is the number of replications, d is the
margin of error, Ỹ is the true mean, and Zα is the value of the standardized variate corresponding to the
level of significance α obtained from the tables.
Sampling Design describes the manner in which sampling units (sample plants) are selected from the
population (ie plot). Three common sampling designs are:
a) simple random sampling- means that each of the N units in the plot is given the same chance of
being selected to compose the n sampling units (n<N). This can be accomplished in many ways:
1. Number all plants in a plot from 1 to N and select at random n numbers using any of the
procedures described earlier. 2. Take a random pair of numbers, with the 1st corresponding to the
width and the 2nd to the length of the plot; the unit at the point of intersection of the random pair
of numbers is then chosen. For example, a sample of six hills (n=6) is to be selected at random
from a potato experimental plot consisting of 10x 20 = 200 hills (N=200). Six pairs of numbers,
41
the 1st number lying between 1 to 10 and the second between 1 to 20 will be chosen from the
table of random numbers:
7,6 2,3 9,15 6,20 3,9 1,10
* * * * * * * * *  * * * * * * * * * * * * * *
* *  * * * * * * * * * * * * * * * * * * * * * random-pair technique for
* * * * * * * *  * * * * * * * * * * * * * * * locating six sample
* * * * * * * * * * * * * * * * * * * * * * * * hills in a plot
* * * * * * * * * * * * * * * * * * * * * * * * consisting of 10 x 20
* * * * * * * * * * * * * * * * * * * * * * *  hills
* * * * *  * * * * * * * * * * * * * * * * * *
* * * * * * * * * * * * * * * * * * * * * * * *
* * * * * * * * * * * * * *  * * * * * * * * *
* * * * * * * * * * * * * * * * * * * * * * * *
b) Multistage sampling- in certain cases the appropriate sampling unit may not coincide with the
unit upon which the measurement of a character is made. For example, in measuring panicle
length in barley the unit of measurement is the panicle. It is however, impractical to use the
panicle as a sampling unit in a simple random sampling scheme since the process would require
the counting and listing of all length of all panicles, a very time consuming task. On the other
hand, it is also not practical to measure the length of all panicles in each sample hill. Thus
sampling designs other than a simple random sampling are necessary. A logical alternative is to
take a simple random sample of n1 hills from each plot, and from each of the selected n1 hills,
take a simple random sample of n2 panicles. The total number of sample panicles, whose length
will be measured, is (n1)(n2). Such sampling procedure is called a two-stage sampling with ‘hill’
as the primary sampling unit and ‘panicle’ as the secondary unit. This method can be easily
extended to three or more stages. In estimating leaf area, for instance, ‘hill’ may be considered as
primary unit, ‘tiller’ as secondary unit, and ‘leaf’ as tertiary unit.
c) Stratified random sampling: here, the plot is 1st subdivided into S subsections and then a simple
random sample of n sampling units is selected from each of these subsection. Such subsections
are referred to as ‘strata’. For example, for barley plots, each consisting of 200 hills, the plot can
be divided 1st into two strata each consisting of 10 x 10 hills. From each stratum, three plants can
then be selected at random. Such stratification increases the precision of the estimate if the
sampling units between strata are more widely different than those within the same stratum. This
is analogous to the concept of blocking in experimental design.
-the technique of stratification can be incorporated with that of multistage sampling, resulting in a
stratified multistage sampling. For example, in estimating the average number of grains per panicle, a
simple random sample of n hills is 1st taken from the plot; and finally a simple random sample of p
panicles is taken from each of the two strata. The total number of sample panicles where the number
of grains will be counted is 2(n)(p). The reason for the stratification here is that the panicles on the
upper portion of a hill are usually bigger and have more grains than those of the lower portion. Thus,
panicles within the same stratum can be expected to be more uniform.
SOIL HETEROGENEITY
Adjacent plots, planted simultaneously to the same genotype, and treated as alike as possible, will
differ in very many characters. The cause for these differences are many, but the most important is
soil heterogeneity.
A Good Experimental Site- to choose an experimental site having minimum soil heterogeneity, the
researcher must be able to identify the features that magnify soil differences, such as: a) Slopes-
fertility gradients are more pronounced in sloping areas, with lower portions more fertile than areas at
the top. Soil nutrients useful to plants are soluble in water and settle in lower areas. An ideal
experimental area should, therefore, have no slope. If this is not available an area with a uniform and
gentle slope is preferred because such areas have predictable fertility gradients that are manageable
through proper blocking. b) Areas used for experiments in previous croppings- different treatments
used in experimental planting usually produce additional soil heterogeneity. Such areas should be
42
avoided if possible, otherwise they should be planted to a uniform variety and fertilized heavily and
uniformly for at least one season before conducting an experiment. Another source of soil
heterogeneity is the presence of unplanted alleys so commonly used in field experiments. Plants
grown in previously unplanted area have been shown to perform better. To avoid additional errors,
unplanted areas should be marked so that the same locations are left as alleys again in succeeding
plantings. c) Graded areas- grading scrapes top soil from elevated portions and dumps it over the
lower areas of the site. This operation, while reducing the slope, results in uneven depth of surface
soil and exposes infertile subsoils. An area that has been graded or has had any kind of soil movement
should be avoided. If this is not possible, it is advisable to conduct a uniformity trial to assess the
pattern of soil heterogeneity so that a suitable remedy can be achieved either by proper blocking or by
appropriate adjustments for the effects of soil heterogeneity in the succeeding crops. d) Presence of
large trees, poles buildings, etc- areas surrounding permanent structures are usually unproductive
because the shade of these tall structures and probably some digging during their construction could
contribute to poor crop performance. Avoid these areas therefore. e) Unproductive site- the area to be
used should be reasonably well suited for the crop to be grown. A productive crop is an important
prerequisite to a successful experiment. Thus an area with poor soil should not be used, unless the
experiment is set up specifically to evaluate such conditions.
Dealing With Soil Heterogeneity: 4 procedures to cope with different types of soil heterogeneity
are► 1. Plot size- two main considerations involved in choosing plot size are practical considerations
and the nature and size of variability. Practical considerations include ease of management in the
field. For variability, the size of experimental error is directly related to soil heterogeneity. While
variability in a given area becomes smaller as plot size becomes larger, the gain in precision decreases
as plot size increases. Also, higher costs are involved when very large plots are used. Hence the plot
size that the experimenter should aim for is one that balances precision and cost. This is referred to as
optimum plot size. 2. Plot shape- the contribution of soil heterogeneity to experimental error results
from differences in soil fertility among plots within a block. The smaller this difference is, the
smaller is the error. Choice of suitable plot shape, therefore, should reduce the differences in soil
productivity from plot to plot within a block and consequently experimental error. This can be done
by orienting plots so that its length runs along the fertility gradient. For areas with a unidirectional
fertility gradient, narrow plots are more advantageous, but for areas with fertile areas in spots, square
or nearly square plots may be better. Square plots are also advisable either when the experimenter
does not know the fertility pattern of the experimental area or when border effects are large. 3. Block
size and shape- are closely related to the size and shape of plots. For a given number of treatments,
the large the plot size, the larger is the size of the block. Thus, the size of block is governed by the
plot size chosen, the number of treatments tested, and the design used. Once these factors are fixed,
only the choice of block shape is left to the researcher. The primary objective in choosing the shape of
blocks is to reduce the differences in productivity levels among plots within a block so that most of
the soil variability in the area is accounted for by variability among blocks. The pattern of soil
heterogeneity in the area is helpful in making this choice. For example, in an area with a
unidirectional gradient, the length of the block should be oriented perpendicular to the direction of the
fertility gradient. Blocks should be kept square or as nearly square as possible if the fertility pattern of
the area is unknown to the researcher or is spotty. 4. Number of replications- number of replications
appropriate for an experiment is affected by a) the inherent variability of the experimental material b)
the experimental design used c) the number of treatments to be tested, and d) the degree of precision
desired.
Since experimental variability is a major factor affecting the number of replications, clearly soil
heterogeneity plays a major role in determining the number of replications in field experiments. In
general, fewer replications are required with uniform soil.
43

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AHS 2307 EXPERIMENTAL DESIGN

The Need For Statistical Evaluation

Characteristics of a Well Planned Experiment

PROCEDURE FOR EXPERIMENTATION

where f= frequency of occurrence of any variate given

Means different, std

R represents number of variates in the sample.

dry weight of 5 plants, Ÿ=3

r is the number of variates in the population

S=2.5 =1.58 g/plant

(Yi)2/r is called the correction term, C. C= (Yi)2/r

S2 = (32+42 + 52 + 22+12 -(3+4+5+2+22)/5)/5 - 1 =2.5

SAMPLING FROM A NORMAL DISTRIBUTION

t DISTRIBUTION AND CONFIDENCE LIMITS

For each sample compute Ÿ, S, SŸ and another statistic, t, where t=(Ÿ-)/SŸ

Ȳ= 24.92308 df = 22 Ȳ= 26.63636

Varieties Replications Yi. Ÿi.

S2w = S21 + S22 /2

Sources of Degrees of Sum of Mean Observed F Tabular F

MSV = SSV/(df)v = 22.5/1 = 22.5

Note that we use totals, not means, in computing SSV.

A POPULATION OF MEAN DIFFERENCES

t TESTS FOR SIGNIFICANCE

Sd = S2y1 + S2y2 =  S21/r1 + S22/r2 = 6.5/5 + 4/5 = 10.5/5 = 2.10 = 1.449

4. Rank the 20 selected numbers from the smallest to largest

cv = 15.1% **= significant at 1%

Correction factor (c.f.) = (grand total)2/total number of observations

ANALYSIS OF VARIANCE: Completely randomized design with an unequal number of

Treatment SS = i.T2i/rI- C.F

RANDOMIZED COMPLETE BLOCK DESIGN

Randomization and layout

Source of Degrees of Sum of Mean Tabular F Block df = total number of blocks - 1 = 4 - 1 = 3

=(30953)2 + (31284)2 + (29846)2 + F = treatment MS/error MS = 239666/110558 = 2.17

Randomization and layout

Step 2: Randomize the row arrangement as follows:

Random number Selection sequence Rank

This plan becomes the final layout of the experiment.

Analysis of Variance: Grain yield of 3 maize hybrids and a check variety

Step 3: outline the analysis of variance:

Step 5: Compute the various sums squares (SS):

Total SS = x2– C.F

Row SS = (row total)2/t – C.F

Error SS = total SS – row SS – column SS – treatment SS

Row and Column Blocking

Variety Any of the basic designs for single-factor

Yield (t/ha) Treatment A two-factor

Block I Block III

Computation steps are: Nitrogen Yield (t/ha)

Step 4; Calculate the various sum of squares as follows:

RANDOMIZATION AND LAYOUT

Random assignment of nitrogen levels to main plots.

Block I Block II Block III

Total SS = x2 – C.F

cv(a) = (error (a) MS/grand mean) * 100 = (141968/5479)*100 = 6.9%

CHI – SQUARE TEST.

X2 = pi=1 (observed  – expected )2 /expected 

Distribution of χ2 for 2, 4, and 6 degrees of freedom

TEST FOR INDEPENDENCE IN A CONTINGENCY TABLE

COMPARISONS AMONG TREATMENT MEANS

DUNCAN’S MULTIPLE RANGE TEST (DMRT)

ACCURACY AND PRECISION

Precision may be improved by:

2. Careful selection of treatments

4. Selection of experimental material