Chapter-III

Material and Method

The present investigation was carried out during rabi season of 2012-2013 at

Vegetable Research Farm, Institute of Agricultural Sciences, Banaras Hindu

University, and Varanasi, which is situated at 25o02΄ North latitude and 83o03΄ East

longitude and at altitude of 128.93 m from sea level. The soil of the experimental

plot was fertile, alluvial loam and was characterized as the type soil of Indo-Gangetic

plains. The place received an annual rainfall of about 1100 mm, major part (88 per

cent) of which was received during July – September months. The mean monthly

meteorological data for the experimental duration is given in appendix-I (Fig. 1-4).

EXPERIMENTAL MATERIAL USED

In the present investigation the experimental material used comprised of 38

genotypes / cultivars of tomato (Solanum lycopersicum L.) obtained from the

NBPGR, New Delhi, Indian Institute of Vegetable Research, Varanasi and from

various other sources and maintained at Department of Genetics and Plant Breeding,

Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221 005. The

genotypes used are listed in the Table-1.

 Material and Method

METHODS

All the 38 genotypes were grown in rabi season of 2012-2013. The crop was planted

on 9th August, 2012. The normal agronomic practices were followed. Fertilizers were

applied at the rate of 120 kg N, 60 kg P 2O5 and 60 kg K2O per hectare, of which

basal application of half of nitrogenous and entire quantities of phosphatic and

potassium fertilizers were given at sowing time. The remaining half amount of

nitrogen was divided into two and was applied after each of two irrigations.Other

crop protection measure were followed as per recommended schedule.

A. Layout Plan

Thirty-eight genotypes were grown in a randomized block design (RBD) with three

replications. Each entry in a replication was represented by two rows, and each row

having ten plants each row measuring 3 meter. Row to row distance was kept at 60

cm and plant to plant distance within the row was maintained at 45 cm.

B. Observations

Five plants were randomly chosen from each entry in each replication and were

tagged for recording detailed observation on yield and yield traits. The following

characters were taken for the present study and the data on different characters were

recorded as described below:

1. Days to first flowering

The numbers of days were counted from the date of transplanting to days to first

flowering.

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 Material and Method

2. Days to 50 per cent flowering

The number of days were counted from the date of transplanting to 50 per cent of

plants flowered.

3. Days to 50 per cent fruiting

The number of days was counted from the date of transplanting to 50% fruiting of

plant population.

4. Number of Primary branches per plant

Total number of branches emerging from the main shoot was counted in each plant

while measuring the plant height and mean value was calculated.

5. Number of Secondary branches per plant

Total number of branches emerging from the primary branches was counted while

data of primary branches were recorded.

6. Plant height (cm)

The plant height was measured from ground level to tip of the plant expressed in

centimetres and mean was computed.

7. Number of clusters per plant

The number of clusters per plant was recorded at the time of harvesting.

8. Number of flowers per cluster

Recorded as average of five random clusters at flowering stage .

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 Material and Method

9. Number of fruits per cluster

Three clusters in each plant were taken at random and the number of fruits in each

cluster was counted. Then the average number of fruits per cluster was calculated.

10. Number of fruits per plant

The total number of marketable fruits harvested from the five plants was counted and

the average number of fruits per plant was calculated.

11. Pericarp thickness (cm)

The fruits were cut at the equatorial plane and the pericarp thickness was measured

with the help of vernier calipers and expressed in centimeter.

12. Number of locules per fruit

The fruits were halved transversely and the locule numbers were counted from the

five fruits. The average was worked out.

13. Total soluble solids content of fruit (TSS)

Tomato juice was collected from red ripe fruits. A drop of juice was placed over the

prism of hand refractometer, with the help of ‘Erma’ (0-33 0 Brix) hand

refractometer and brix value was noted in per cent at room temperature.

14. Seed Index

Bold and healthy hundred seeds were randomly counted and was weighed on

electronic balance and recorded in gram.

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 Material and Method

15. Average fruit weight (gm)

The total number of marketable fruits were weighed and the average fruit weight was

worked out and expressed in grams (gm).

16. Fruit yield per plant

The weight of fruits from each picking was recorded from the five labeled plants of

each experimental plots. Total yield per plant was worked out by adding yield of all

harvests and was expressed in kilogram (kg) per plant.

17. Juice-pulp ratio.

Randomly 10 fruits were selected and their weight is measured and recorded. Then

juice was properly extracted from the fruit and remaining pulp was weigh.

Difference between the fruit weight and pulp weight give the weight of juice. From

the data juice: pulp ratio was calculated.

18. Fruit shape index

It is measured by both polar diameter and equatorial diameter of fruit and expressed

in centimetres. Fruit polar diameter was measured from stalk end to blossom end by

using vernier calipers. Fruit equatorial diameter was measured from fruit breadth at

highest bulged portion of the fruit by using vernier calipers.

19. Carotene content (mg/100g)

It was estimated by colorimetric method described by Grag et a.l (2008)

with the help of UV-Vis spectrophotometer by taking optical density at 450nm

37
 Material and Method

wavelength. Same procedure followed for the estimation of carotene content except

the wavelength used was 450 nm.

20. Lycopene content (mg/100g)

Same procedure followed for the estimation of carotene content except the

wavelength used was 505 nm.

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 Material and Method

STATISTICAL ANALYSIS

ANALYSIS OF VARIANCE

The mean value of five plants from each entry in each replication was used for the

detailed statistical analysis. The data were analysed by Analysis of Variance

technique of Randomized Block Design given by Ostle (1966).

Treatments
Replication
Total
s
1 2 3 4 T

1 X11 X12 X13 X14 X1t R1

2 X21 X22 X23 X24 X2t R2

3 X31 X32 X33 X34 X3t R3

4 X41 X42 X43 X44 X4t R4

r Xr1 Xr2 Xr3 Xr4 Xrt Rr

Total T1 T2 T3 T4 Tt G.T.

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 Material and Method

The analysis of variance in a single environment is based on the mathematical

model:

Yij = μ + gi + bj + eij

Where,

Yij = the mean performance of the ith genotype in jth block

μ = General Mean

gi = the effect of ith genotype

bj = the effect of jth block

eij = the environmental effect specific to the i th genotype in the jth

block

On the basis of the above model, the analysis of variance was set up as follows:

Grand Total
Grand Mean =
Where, N

N = number of treatments (t) x replications (r)

Replication Number

Correction Factor (C.F.) (Grand Total)2

= N

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 Material and Method

r t
∑ ∑ X ij −C . F .
i=1 j=1
Total Sum of Square (T.S.S.) =

Where,

Xij = Observation corresponding to ith row and jth column

Sum of Square of
Treatment Totals
Treatment Sum of Square (S.S.T.) = - C.F.
Replication Number (r)

Sum of Square of
Replication Totals
Replication Sum of Square (S.S.R.) = - C.F.
Treatment Number (t)

Error Sum of Square (S.S.E.) = T.S.S. – (S.S.T. + S.S.R.)

Mean squares (M.S.) were obtained by dividing the sum of squares with the

respective degrees of freedom.

Treatment M.S.
‘F’ (Variance Ratio) =
Error M.S.

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 Material and Method

Analysis of Variance Table (ANOVA)

Source of Expected
d.f. S.S. M.S. ‘F’ Value
Variation M.S.

S .S . R.
M R=
Replication (r – 1) S.S.R. (r−1 ) σ2e + gσ2r

S . S .T . MT
M T=
Treatment (t – 1) S.S.T. (t−1) σe+rσg
2 2
ME

S. S.E.
ME=
Error (r – 1) (t – 1) S.S.E. (r−1 )(t−1 ) σ e
2

Total (rt – 1) T.S.S.

Where,

r = number of replications

t = number of treatments (i.e., genotypes/ cultivars / strains)

σ2g = the genotypic variance

σ2e = the error variance (or error M.S.) = Environmental Variance

Genotypic and Phenotypic Variance:

Error Variance, σ2e = ME

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 Material and Method

M T −M E
σ 2g =
r
Genotypic Variance,

Phenotypic Variance, σ2p = σ2g + σ2e

The significance of selections was tested by comparing the calculated ‘F’ value with

the table value of ‘F’ at 5 per cent level of significance for the corresponding degree

of freedoms. Where ‘F’ test was observed to be significant, comparisons were further

extended by testing difference of any two selections against critical difference (C.D.)

at 5 per cent level of significance (i.e., α = 0.05).

PARAMETERS OF VARIABILITY

Mean:

The mean value of each character was determined by dividing the total number by

corresponding number of observations.

N
∑ Xi
X= i=1
N
Where,

X
= mean,

N
∑ Xi
i=1
= Sum of all observations

N = Number of observations

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Material and Method

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 Material and Method

Range:

The lowest and highest values for each character were taken as the range.

Standard Error of Difference Between Two Means [S.E.d.(M)]:

S.E.d.(M) was calculated with the help of error mean square from ANOVA.

S .E .d.( M )=
√ 2 M .S .E .
r

Where,

r = Number of replications

M.S.E. = Mean sum of square due to error

Critical Difference (C.D.):

Significant ‘F’ value indicates that there is significant difference among the

treatments i.e., the genotypes. Then we have to further search for the genotypes

which are significant difference between them for the character under study. For this

purpose we have to calculate a least significant difference which is also known as

critical difference, with which the difference between all the treatment combinations

have to compare. Any genotype combination showing difference more than or equal

to critical difference will be significantly different in their performance for the

character under study.

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 Material and Method

Critical Difference (C.D.) =

√ 2 M . S .E .
r
x ‘t’at α = 0.05; at error d.f.

Coefficient of Variation:

σP
Phenotypic Coefficient of Variation =
(PCV) X x 100

Phenotypic and genotypic coefficients of variability were calculated by the method

suggested by Burton and Devane (1953):

σg
Genotypic Coefficient of Variation =
(GCV) X x 100

Where,

Ϭp = Phenotypic standard deviation

Ϭg = Genotypic standard deviation

X = General mean

Heritability:

It was calculated by the formula given by Allard (1960) which is as below:

2
σg
H= 2
σp

Where,

H = Heritability in broad sense

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 Material and Method

σ2g = Genotypic variance

σ2p = Phenotypic variance

Genetic Advance:

The genetic advance, i.e., expected genetic gain resulting from selecting five per cent

superior plants was estimated by the following formula suggested by Allard (1960):

√ σ 2p
Genetic Advance (Expected) = H x xk

Where,

H = Heritability coefficient

√ σ 2p = Phenotypic standard deviation

K = Selection differential in standard units which is 2.06 for

5% selection intensity.

Genetic advance as percentage of mean was calculated by the following formula:

( GAX )×100
Genetic Advance as Percentage of Mean =

Where,

GA = Expected genetic advance

X
= General mean of the character in the population

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Material and Method

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 Material and Method

CORRELATION COEFFICIENT

The genotypic and phenotypic correlation coefficients were calculated according to

the formula suggested by Searle (1961).

Cov (X1X2)
r(X1X2) =
√ V ( X 1 )⋅V ( X 2 )
Where,

r (X1X2) is the correlation between characters X1 and X2

Cov X1X2 is the covariance between X1 and X2

V(X1) is the variance of X1

V(X2) is the variance of X2

In phenotypic correlation coefficients, phenotypic covariance and variance are

considered for calculation whereas in case of genotypic correlation coefficient,

genotypic values are taken. To test the significance of correlation coefficients, the

estimated values were compared with the table value (statistical table by Fisher and

Yates, 1963) at n-2 degrees of freedom (where n denotes the number of genotypes

tested) at 5% and 1% level of significance, respectively.

PATH COEFFICIENT ANALYSIS

Path coefficient analysis was done to partition the total correlation into direct and

indirect effects due to the dependent variable. Sewall Wright (1921) suggested this

analysis and it was further elaborated by Dewey and Lu (1959).

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 Material and Method

Path coefficient is the ratio of standard deviation of the effect due to a given cause to

the total standard deviation of the effect, i.e., if yield per plant (Y) is the function of

the causal factor X1, then path co-efficient for the path from causal factor X1 to the

effect Y is σX1/ σY.

In other word, it is a standardized partial regression coefficient, which individually

provides a measure of direct effect of the causal factor or independent variables on

the effect dependent variable. These permit partitioning of the correlation between

the causal factor and the effect of variable into components of direct and indirect

effects and this makes the association of causal factor with the effect of variable

more clear.

Here yield per plant (Y) was taken as ‘effect’ the other characters X 1, X2, X3,……Xn

were related to seed yield as causal factors.

The path coefficients were obtained by solving a set of simultaneous equations of the

form:

r X Y =P X Y +r X X P X Y +r X X P X Y +. .. .. . .. .. .. . .. .. . .. .. . .. .. . .+r X Xn PX Y
1 1 1 2 2 1 3 3 1 n

r X Y =r X 2 X1 P X1 Y + PX 2 Y + r X 2 X 3 P X 3 Y +. .. .. . .. .. . .. .. . .. .. .. . .. .. . .+ r X2 X n P X n Y
2

r X Y =r X n X 1 P X 1 Y +r X n X2 P X2 Y +r Xn X 3 P X 3 Y +. . .. .. . .. .. . .. .. .. . .. .. . .. .. .+ P X n Y
n

Where,

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 Material and Method

rX Y rX
1 nY

to denotes coefficient of correlation between independent

characters X1 to Xn and dependent character Y.

rX rX
1 X2 n−1 X n

to denotes coefficient of correlation between all possible

combinations of independent characters.

PX Y PX Y
1 n

to denotes direct effects of character X1 to Xn on Y.

The solutions for path coefficients, direct and indirect effects of the causal factor

were calculated as per the values of right hand side of equation.

The residual factor (undefined factor), i.e. the variation in yield unaccounted for

causal effects under consideration (PRY) or the path values for residual effect was

calculated as follows:

√ 1−R2
Residual factor (PRY) =

∑ PX Y r X Y
i i
i=1
Where, R2 =

R2 is the coefficient of determination and is the amount of variation in yield and that

can be accounted for the yield component trait. Here, path coefficient at phenotypic

level was calculated using phenotypic correlation coefficient.

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