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When you run an ANOVA (Analysis of Variance) test and get a significant result, that means at least one of the groups tested differs from the other groups. However, you can’t tell from the ANOVA test which group differs. In order to address this, Fisher developed the least significant difference test in 1935, which is only used when the ANOVA F omnibus returns a significant result. The LSD calculates the smallest significant between two means as if a t-test had been run on those two means. This enables you to make direct comparisons between two means from two groups. Any difference larger than the LSD is considered a significant result.

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OF TREATMENT MEANS

Comparing means of

different treatments i.e.

which two treatments are

different significantly?

Comparison of means is

necessary only when there is

significant difference among

treatment means as indicated

by ANOVA

H

o

: u

1

= u

2

= u

3

= ……= u

t

When H

o

is rejected, we conclude that

the means differ significantly (at least 2

means differ significantly), but does not

specify which pairs of treatment means

differ. LSD, DNMRT and HSD can be

applied.

LSD (Least significant

difference) Test

Designed by Fisher

It’s a t-test.

More suitable for

comparing paired means.

d

s

Y Y

t

2 1

Therefore:

If the observed difference between the two means exceeds LSD the

difference is declared as significant, thus:

d

s t LSD .

r

MSE

s

d

2

Where:

d

s t Y Y .

2 1

Arrange data according to treatments and replications:

Treatment RI RII RIII RIV Total Mean

V1 47 52 62 51 212 53

V2 50 54 67 57 228 57

V3 57 53 69 57 236 59

V4 54 65 74 59 252 63

Total 208 224 272 224 928

Mean 52 56 68 56

ANOVA Table

Source

Variation

df SS MS F F

table

(α=0.05)

Total 15 854

Rep. 3 576 192 24.7* 3.86

Treatment 3 208 69.3 8.9* 3.86

Error 9 70 7.78

Calculation :

Obtain value of t from table at df = dfe = 9 and

α=0.05, t = 2.262

therefore LSD = 2.262 x 1.972

= 4.46 t/ha

d

s t LSD .

r MSE s

d

/ 2

972 . 1 4 / 78 . 7 2

d

s

Compare difference between treatment means with LSD:

Larger significant

Smaller not significant

Variety Var1 Var2 Var3 Var4

Mean 53 57 59 63

V1 vs V4 = 53 – 63 = 10 > 4.46 significant difference

V1 vs V3 = 53 – 59 = 6 > 4.46 significant difference

V1 vs V2 = 53 – 57 = 4 < 4.46 no significant difference

V2 vs V4 = 57 – 63 = 6 > 4.46 significant difference

V2 vs V3 = 57 – 59 = 2 < 4.46 no significant difference

V3 vs V4 = 59 – 63 = 4 < 4.46 no significant difference

A CRD experiment on nitrogen content of red clover plants

innoculated with combination cultures of Rhizobium trifolii

strains and Rhizobium meliloti strains with 6 treatments

and 5 replications results the following:

a b c d e f

19.4 17.7 17.0 20.7 14.3 17.3

32.6 24.8 19.4 21.0 14.4 19.4

27.0 27.9 9.1 20.5 11.8 19.1

32.1 25.2 11.9 18.8 11.6 16.9

33.0 24.3 15.8 18.6 14.2 20.8

Exercise:

Test the different combinations using LSD.

a b c d e f

Y

.j

R1 19.4 17.7 17.0 20.7 14.3 17.3 106.4

R2 32.6 24.8 19.4 21.0 14.4 19.4 131.6

R3 27.0 27.9 9.1 20.5 11.8 19.1 115.4

R4 32.1 25.2 11.9 18.8 11.6 16.9 116.5

R5 33.0 24.3 15.8 18.6 14.2 20.8 126.7

Y

i.

144.1 119.9 73.2 99.6 66.3 93.5 596.6

Y 28.8 24.0 14.6 19.9 13.3 18.7

Calculation:

Source

Variation

df SS MS F F

table

(α=0.05/0.01)

Total 29 1129.98

Treatments 5 847.05 169.41 14.37** 2.62/

3.90

Error 24 282.93 11.79

Calculation :

CF =

11864.39

Obtain value of t from table at df = dfe = 24,

α=0.05, t = 2.064

therefore LSD = 2.064 x 2.172

= 4.483 mg N

d

s t LSD .

r MSE s

d

/ 2

172 . 2 5 / 79 . 11 2

d

s

Compare difference between treatment

means with LSD= 4.483 mg N :

a vs e = 28.8 – 13.3 = 15.5 > 4.483 significant difference

a vs c = 28.8 – 14.6 = 14.2 > 4.483 significant difference

a vs f = 28.8 – 18.7 = 10.1 > 4.483 significant difference

a vs d = 28.8 – 19.9 = 8.9 > 4.483 significant difference

a vs b = 28.8 – 24.0 = 4.8 > 4.483 significant difference

b vs e = 24.0 – 13.3 = 10.7 > 4.483 significant difference

b vs c = 24.0 – 14.6 = 9.4 > 4.483 significant difference

b vs f = 24.0 – 18.7 = 5.3 > 4.483 significant difference

b vs d = 24.0 – 19.9 = 4.1 < 4.483 insignificant difference

Contd .......

cultures

e c f d b a

mean 13.3 14.6 18.7 19.9 24.0 28.8

d vs e = 19.9 – 13.3 = 6.6 > 4.483 significant difference

d vs c = 19.9 – 14.6 = 5.3 > 4.483 significant difference

d vs f = 19.9 – 18.7 = 1.2 < 4.483 insignificant difference

f vs e = 18.7 – 13.3 = 5.3 > 4.483 significant difference

f vs c = 18.7 – 14.6 = 2.1 < 4.483 insignificant difference

c vs e = 14.6 – 13.3 = 1.3 < 4.483 significant difference

HSD (Honestly Significant

Difference) - Tukey

Tukey (1949) proposed a multiple

comparison procedure where all pairs

of treatment means can be compared

Like LSD, a single value is used for

judging significant difference, called

HSD

HSD is higher than LSD, thus more

conservative.

Obtain Q from table refering to df = dfe = 24,

k = 6, α=0.05, where k = number of treatments i.e

Q = 4.37.

Thus

r MSE Q HSD /

mgN HSD 73 . 6 54 . 1 37 . 4 5 / 79 . 11 37 . 4

Compare difference between treatment

means with HSD = 6.75 mg N :

a vs e = 28.8 – 13.3 = 15.5 > 6.73 significant difference

a vs c = 28.8 – 14.6 = 14.2 > 6.73 significant difference

a vs f = 28.8 – 18.7 = 10.1 > 6.73 significant difference

a vs d = 28.8 – 19.9 = 8.9 > 6.73 significant difference

a vs b = 28.8 – 24.0 = 4.8 < 6.73 insignificant difference

b vs e = 24.0 – 13.3 = 10.7 > 6.73 significant difference

b vs c = 24.0 – 14.6 = 9.4 > 6.73 significant difference

b vs f = 24.0 – 18.7 = 5.3 < 6.73 insignificant difference

b vs d = 24.0 – 19.9 = 4.1 < 6.73 insignificant difference

Contd .......

cultures

e c f d b a

mean 13.3 14.6 18.7 19.9 24.0 28.8

d vs e = 19.9 – 13.3 = 6.6 < 6.73 insignificant difference

d vs c = 19.9 – 14.6 = 5.3 < 6.73 insignificant difference

d vs f = 19.9 – 18.7 = 1.2 < 6.73 insignificant difference

f vs e = 18.7 – 13.3 = 5.3 < 6.73 insignificant difference

f vs c = 18.7 – 14.6 = 2.1 < 6.73 insignificant difference

c vs e = 14.6 – 13.3 = 1.3 < 6.73 insignificant difference

Treatments Mean

a 28.8a

b 24.0ab

C 14.6c

d 19.9bc

e 13.3c

f 18.7bc

HSD = 6.73 mg N

Treatments Mean

a 28.8a

b 24.0b

c 14.6c

d 19.9bd

e 13.3c

f 18.7cd

LSD = 4.48 mg N

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