Matedals and Methods
3.12 Data recording
Data were recorded from each pot based on different agromorphogenic, physiological and nutritional traits. Different steps of data collection are presented in Plate S. Data were recorded in respect of' following parameters:
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Plate 4. Intercultural operations. A) Watering in seedhed, B) Pesticide application in tomato plants
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PlateS. Different types of data recording. A) Size and fruit weight using digital balance, 0) Extraction of vitamin- C by Oxidation Reduction Titration Method, C) Estimation of lycopene content using T60 UV-Visible Spectrophotometer, D) Estimation of brix (%) using Portable Refractometer, E) Determination of chlorophyll content SPAD-502 plus Portable ChLorophyll meter, F) Estimation of Na and K content using
3.12.1 Agromorphogenic traits
Different agrornorphogenic traits related to yield and its contributing characters were recorded viz., days to first flowering plant height. number of' clusters per plant. days to maturity, number of fruits per cluster. number of fruits per plant. average fruit length (mm). average fruit diameter (mm), average fruit
weight per plant (g) and yield per plant (kg).
3.12.1.1 Days to first flowering
The number of days to first flowering was counted from the date of tomato seedlings transplanting to date of first flowering.
3.12.1.2 Plant height (cm)
Plant height of each plant at mature stage measured in em using meter scale and mean was calculated.
3.12.1.3 Number of clusters per plant
The number of clusters per plant was recorded at the time of harvesting.
3.12.1.4 l)ays to maturity
The number of clays to maturity was counted from the date of tomato genotypes transplanting to date of first harvesting.
3.12.1.5 Number of fruits per cluster
All fruits per cluster were recorded and then the average number of fruits per cluster was calculated by randomly selecting three clusters.
3.12.1.6 Number of fruits per plant
The total number of marketable fruits harvested from each plant was recorded.
3.12.1.7 Average fruit length and diameter (mm)
Fruit length and diameter were measured using Digital Caliper-S 15 (DC-S 15) in millimeter (mm). Mean was calculated for each treatment.
3.12.1.8 Average fruit weight per plant (g)
Fruit weight was measured by electric precision balance. Average fruit weight per plant was recorded by randomly selecting 5 fruits per plant and mean value was calculated.
3.12.1.9 Yield per plant
Yield per plant was recorded from all hanests of each plant and expressed in kilogram (kg) per plant.
3.12.2 Physiological traits
Data related to different physiological traits such as chlorophyll content (%).
Na content (%). and K content (%) were recorded.
3.12.2.1 Measuring of chlorophyll content
Leaf chlorophyll content was measured by using SPAI)-502 plus Portable Chlorophyll meter. The chlorophyll content was measured at 92 DAT from
lour different portion of the leaf and then averaged for analysis.
3.12.2.2 Determination of Na' and K content
Oven-dircd (70°C) tomato plants shoot samples were ground in a W'iley Hammer Mill. passed through 40 nicsh screens. mixed well and stored in plastic vials. The ground plant samples were digested by Micro-Kjeldahl method (Thomas and Nambisan, 1999). One grain oven-dried tomato plant shoot samples were taken in kjeldahl flasks. About 15 ml. of diacidic mixture (HNO3: 60% ITClOi 2:1) were taken in a digestion tube and lefl to stand for 20 minutes and then transferred to digestion block and continued heating at 100°C. The temperature was increased to 365°C gradually to prevent frothing (50°C steps) and left to digest until yellowish color of the solution turned to whitish color. 1hen the digestion tubes were removed from the heating source and allowed to cool to room temperature. About 40 inL of de-ionised water was carefully added to the digestion tubes and the contents l9ltered through Whatman no.40 filter paper into a 100 mL volumetric flask and the volume up to the mark with de-ionised water. The samples were stored at room temperature in clearly marked containers. Content of Na' and K were determined by Flame Photometer.
3.12.3 Nutritional traits
Data were recorded on the basis of different nutritional traits using ripe thiitcs viz.. Brix (%). Vitamin-C content (m!100 ) and Lycopene content (1110i]00 g).
3.12.3.1 Determination of Brix percentage
Brix percentages was measured by Portable Refractometer (ERMA. Tokyo.
Japan) at room temperature. Single fruit was blended and juice was collected to measure brix percentage.
3.12.3.2 Determination of Vitamin-C
Vitamin-C was measured by Oxidation Reduction Titration Method (Tee ci at.
1988). Single fruit was blend and tomato extract was filtrated by Whatrnan No.1 filter paper. It was then mixed with 3% nietaphosphoric acid solution.
The titration was conducted in presence of glacial acetic acid and meiaphosphorie acid to inhibit aerobic oxidation with dye solution (2. 6- dichlorophenol indophenol). The solution was titrated with dye. The observations mean will give the amount of dye required to oxidize definite amount of L-ascorbic acid solution of unknown concentration. using 1.-ascorbic acid as known sample.
3.12.3.3 Determination of Lycopeue content
Absorption determination for lycopene content was estimated following the method of Alda ci at. (2009) by using T60 UV-Visihle Spectrophotometer.
Lcopene in the tomato was extracied using hexane ethanol acetone (2:1:1) mixture. One gram juice of the each sample was homogenized with 25 ml of hexane : ethanol : acetone. which were then placed on the orbital shaker for 30 mm.. adding 10 ml distilled water and was continued agitation tbr another two mm. The solution was then left to separate into distinct polar and non- polar layers. The absorbance was measured at 472 nm and 502 mn, using hexane as a blank. The lycopene concentration was calculated using its sped tic extinction
coefficient (IT 1%, 1 cm) of 3450 in hexane at 472 nni and 3150 at 502 nrn. The lycopene concentration was exprcsscd as mg/I00 gproduct.
E 20
At X = 472 nni : lycopene content (mg/100 g) = ______ ____
3.45 In
ii 20
At ?. = 502 run lycopene content (mg /100 g) = _____
3.15 m
Where.
m = the weight of the product (g) E = extinction coefficient
3.13 Statistical analysis
Collected data were statistically analyzed using MSTAT-C computer package program. Mean for every treatments were calculated and analysis of variance for each one of characters was performed by F-test (Variance Ratio). Different between treatments was assessed by Least Siiflcant Difference (LSD) test at 5% Level of significance (Gomez and Gomez. 1984).