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The Effects of Planting Distances and Different Stages of Maturity on the Quality of Three Cultivars of Tomatoes ( Lycopersicon esculentum Mill)

Atefeh TABASI

1

*, Hossein NEMATI

1

, Mohammad AKBARI

2

1Ferdowsi University of Mashhad, Faculty of Agriculture, Department of Horticulture, Mashhad, Iran; [email protected] (*corresponding author)

2Rasht University of Guilan, College of Agriculture, Department of Horticulture, Guilan, Iran

Abstract

In this investigation the effects of cultivar, row spacing and different stages of maturity on qualitative characteristics of tomato (ascorbic acid, total soluble solids (TSS), β-carotene and lycopene) have been evaluated. Experiment was performed by factorial analysis with 3 replicates in completely randomized design (CRD). First treatment was three cultivars of tomato, second treatment was four planting distances and third treatment was different stages of maturity. The results showed that all treatments had significant influence on the levels of ascorbic acid, soluble solids, β-carotene and lycopene. Generally, wider spacing and deep red fruits had the highest quality.

Therefore, choosing appropriate cultivars, special planting distances and suitable stage of maturity can increase fruit quality of tomato.

Keywords: ascorbic acid, carotenoid, lycopene, soluble solids, spacing

Introduction

Lycopen, a participant in the carotenoid family along with a pigment which attributes to the red color of to- matoes, is a leading factor to its health promoting ability (Tonucci et al., 1995; Fanasca et al., 2007). Many studies have shown that tomatoes can help decrease the chance of getting lung cancer, because lycopene acts as antioxidant (Young et al., 1993). Qualitative factors of tomato (flavor, color, soluble solids and nutrition value) were affected by cultivar, weather, condition storage, fruit maturity and cul- tivation methods (Gould, 1983). Changes in tomato fruit ripening occur within the plastids after the disappearance of chlorophyll. The two major groups of pigments found in tomato fruit are chlorophyll and carotenoids. The most noticeable change during ripening is the remarkable in- crease in the carotenoid content of the fruit (Laval-Martin et al., 1975). During ripening the chlorophyll concentra- tion decreases while carotenoids, especially lycopene, ac- cumulate in the fruit (Laval-Martin et al., 1975). Accord- ing to Brandt et al. (2003) higher lycopene content was observed in tomato harvested in glasshouse-grown than in field-grown at different harvesting times. As suggested by Luh and Daoud (1968) the amount of lycopene in dif- ferent cultivars reveals a considerable difference. The light intensity is influential in the biosynthesis of carotenoids and development of fruit color (Shiraghinge et al., 2010).

Ascorbic acid, total soluble solids (TSS) and contents are commonly considered as fruit quality determining prop- erties in tomato. Soluble solids include mainly the sugars such as glucose, fructose and sucrose. In tomato fruit, or-

ganic acids with sugars make a major contribution to the taste of the fruit. Most variation in flavor can be related to differences in the sugars and acids contents of the fruits.

Bradly (1964) and Mauz (1966) showed that there is a fundamental difference in acidity between tomato culti- vars. Although the cultivar has a dominant influence on the quality determinant properties, the environment in which it grows also has a significant impact on quality characters (Purseglove et al., 1986). As shown by Malews- ki and Markakis (1971) ascorbic acid content decreases with ripening thus conflicting results could have been due to differences in ripeness at time of analysis.

Spacing is one of the important factors, which influence the quality of tomato fruit. Many experiments have been conducted on the effect of spacing on growth and quality of tomato. Tanaka and Komochi (1982) studied the rela- tionship between plant density and topping on the growth and quality of tomato in greenhouse. They reported with increasing plant density, leaf size, stem diameter, weight of leaves and stem per plant decreased and flowering as well as ripening was delayed up to 7 days and yield per unit area increased. Srinivas and Hegde (1984) demonstrated that increased spacing increased dry matter production and quality attributes of tomato. A positive correlation was reported between plant density and yield and negative correlation between plant density and quality of fruits, so that the highest fruit quality was recorded in lowest plant density (Manchanda and Bhopal Singh, 1988). Gaye et al.

(1992) reported that quality attributes of tomato declined with increasing plant density (Davies and Hobson, 1981;

Baxter et al., 2005). Srinivasa et al. (1988) observed that Received 16 May 2013; accepted 24 June 2013

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based on the absorbance at 451 nm with an extinction co- efficient of 2505 (Goodwin, 1955).

Lycopene analysis

The tomato samples (approximately 500 mg) were ground in a mortar in liquid nitrogen, followed by grind- ing in 4 ml of acetone with 50 mg CaCO3. After centrifu- gation, pellets were re-extracted with 4 ml of acetone, 2 ml of hexane and 5 of ml acetone: hexane (4:1), successively.

The supernatants were combined, measured and filtered through 0.2 µm nylon syringe filters into HPLC vials. All processes were performed as much as possible under sub- dued or safe light and a nitrogen atmosphere. For HPLC analysis a Spectra Physics SP8800 pump system was used equipped with a Spectra Physics Spectra 100 UV-Vis de- tector (Spectra Physics, Mountain View, CA, USA). For system processing and data acquisition a computer system with WINner on Windows software was used (Thermo Separation Products, Wirral, UK). Sample injections were carried out by means of a Waters 717 (Milford, MA, USA) autosampler equipped with a cooler. Separation of the pigments was carried out according to Gilmore and Ya- mamoto (1991) using a non-endcapped Allsphere ODS-1 HPLC column (4.6 mm × 250 mm, 5 µm particle size) preceded to a ODS-1guard column (Alltech Associates).

Stainless steel column frit/insert material was replaced by a Peek Alloyed with Teflon (PAT) column frit/insert. The column temperature was 30°C. The mobile phase con- sisted of acetonitrile, methanol and tris buffer 0.1 M (pH 8.0). The mobile phase contained 0.1% (w/v) butylated hy- droxytoluene. The flow rate was 1 ml min−1, sample injec- tion volume was 20 µl and spectrophotometric detection was performed at 445 nm. The concentrations of pigment standard stock solutions were determined spectrophoto- metrically using published absorbance coefficients (Kon- ings and Roomans, 1997).

Statistical analysis

Effect of treatments were verified based on ANOVA using Excel program and the means were compared using LSD test at 5% level.

Results and discussion Ascorbic acid

The experiment results demonstrated that there was a vital difference between three cultivars, planting distances and different stages of fruit ripening in ascorbic acid con- tent. So the highest content of ascorbic acid was related to deep red fruits in 49 cm of planting distance (Tab. 2). As shown by Sing and Dillon (1986) the ascorbic acid and other acidity content of tomato juice was increased with maturity stages and reached the peak and thereafter start- ed to decrease Singh and Dillon (1986). Also our results are similar to Mauz (1966) who reported that increasing light in to the plant leads to raised ascorbic acid. It can wider row spacing gave significantly more fruit quality

than narrow spacing in tomato cultivars.

Optimum plant spacing may help in proper utilization of land and for obtaining good quality fruits. On the other hand, stem pruning also may influence on the production of yield. Therefore, the present study was undertaken to investigate the effect of optimum plant density and differ- ent stages of maturity for higher yield and better quality of tomato.

Material and methods

Plant materials and treatments

In this study three cultivars of tomato (‘Super A’, ‘1449’, and ‘Nemark 49’) grown in greenhouses of (Ferdowsi University of Mashhad in Iran). Each tomato cultivar was planting in four distances (25 cm, 33 cm, 41 cm and 49 cm). These planting distances are common in this area The purpose of these intervals, was Investigate the effect of light and environmental conditions on fruit quality. Fruit sample from four maturity stages according Huerres Perez (1987) were gathered randomly by hand in three repeti- tions. Stages of maturity were classified as follows:

Mature green: fruits are mature and entirely light to dark green.

Pink: pink or red color ranges between 10 to 30%.

Red: red color is more than 60% but less than 90%.

Deep-red: red color exceeds 90% (Yamaguchi, 1983).

Traits measured were included soluble solid, ascorbic acid, β-carotene and lycopene at four different stages of fruit ripening.

Soluble solids analysis

Total soluble solid contents were determined by ex- tracting and mixing one drop of juice from each fruit into a refractometer (ERMA, TOKYO).

Ascorbic acid analysis

Ascorbic acid was measured according to the method of AOAC (2005). In order to measure the ascorbic acid, 10 cm3 from tomato juice was mixed with 20cc distilled water and then 2 cc from (1%) one-percent soluble starch was added. Then ascorbic acid was determined by titration of 10 ml filtrated juice which contained potassium iodide (KI). In fact, it was based on mg ascorbic acid per100 g FW. Ascorbic acid content was calculated using the fol- lowing formula (AOAC, 2005).

β - Carotene analysis

The β-carotene was analyzed by a modified AOAC method (AOAC, 1970) and was extracted with an ace- tone-hexane mixture. The MgO-Hyflo Supercel chromato- graphic column was washed with 0.5% acetone in hexane so that isomerization of carotenoids would be minimized (Wiseman et al., 1952). β-carotene was eluted with 5%

acetone in hexane. The concentration of β-carotene was

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be concluded that in 49 cm of row spacing, more light to the crown of the plant was penetrated, which results in increased fruit ascorbic acid. Mahendran and Bandara (2000) reported that moisture stress reduced the ascorbic acid content of tomato fruits when the stress was imposed during the fruit ripening stage. It can be concluded that close spacing retains soil moisture. Also they stated a re- duction in the D-glucose synthesis would have occurred during the period of stress, which in turn may have re- duced the synthesis of ascorbic acid. Moisture stress may have reduced the substrate concentration for ascorbic acid synthesis. Reduction in the substrate may possibly be due to reduced photosynthetic rate (Mahendran and Bandara 2000).

Total soluble solids (TSS)

The study results revealed that there was a significant difference between three cultivars, row spacing and dif- ferent stages of fruit ripening in soluble solids. The most soluble solids were obtained from red fruits of ‘Super A’

cultivar in the 49 cm of plant-spacing. Fernando and Alis- dair (2006) suggested more space of planting distances in- creased photosynthesis which results in increased soluble solids. It should be noted that there are a lot of light pen- etrated in wider spacing in comparison with close spacing.

Purseglove et al. (1986) reported that fruit ripening had a major impact on quality attribute and raised the dry mat- ter of fruit and soluble solids. These results are in close agreements with our findings (Tab. 1).

β –Carotene

The results of this experiment showed that cultivar, row spacing and different stages of fruit ripening had central effects on carotenoied level, so that the highest β-carotene content belonged to ‘Nemark 49’ cultivar in 49 cm of planting distance of pink fruits (Tab. 3).

Young et al. (1993) mentioned that there is a promi- nent difference between cultivars. They also expressed that presence amount of carotenoid in some cultivars and their absence in the other varieties is due to differences in their genes controlling biosynthesis. In addition, they reported that light density is a major factor to improve carotenoid Chlorophyll content. Bradly (1964) stated that fruits re- ceiving more light have higher levels of carotenoid. So, it can be concluded that increasing row spacing, revealed a better penetration of light to the crown of the plant that leads to β- carotene content increasing.

Lycopene

The results demonstrated that cultivar, row spacing and different stages of ripening fruit had an essential difference on lycopene level. The results of our investigation estab- lished that the content of lycopene in all the investigated tomatoes during fruit ripening significantly increased. The lowest concentration of lycopene (32 mg 100 g-1) was re- corded in the green fruits of the cultivar ‘1449’, in 25 cm of row spacing and the highest concentration of lycopene (434 mg 100 g-1) was measured in completely ripened to- matoes of the cultivar ‘Nemark 49’ in 41 cm of row spac- ing (Tab. 4). Muhammad and Singh (2007) reported that Tab. 1. Comparison of interaction between variety, row spacing and different stages of maturity for TSS

Different stages green pink red deep red

Planting distances 25 33 41 49 25 33 41 49 25 33 41 49 25 33 41 49

Cultivar

‘Super A’ 2.2l 2.3k 2.7j 2.8i 3.0h 3.1g 3.1g 3.2f 3.45e 3.46e 3.68d 3.7d 4b 3.9c 4.38a 4. 9a

‘1449’ 2.2k 2.65j 2.65j 2.7j 3.1i 3.35h 3.5g 3.46g 3.68f 3.7f 3.98e 4e 4.3d 4.5c 4.7b 4.4a

‘Nemark 49’ 2.6i 2.3k 2.4j 2.6i 2.76h 2.8h 2.98g 3.0g 3.13f 3.2f 3.36e 3.4e 3.6d 3.7c 3.8b 4.4a Means with similar letters in each table are not significantly different by LSD multiple range test (p< 0.05)

Tab. 2. Comparison of interaction between variety, row spacing and different stages of maturity for ascorbic acid

Different stages green pink red deep red

Planting distances 25 33 41 49 25 33 41 49 25 33 41 49 25 33 41 49

Cultivar

‘Super A’ 11.86i 12.70h 14.46g 14.20g 17.03f 17.26f 20.9ds 20.3de 23.8b 23.2a 27.46a 27.10a 20.10e 20.86d 22.33c 22.40c

‘1449’ 11.73j 12.40i 14.66h 14.06h 16.30g 17.63f 20.3de 19.9d 23.1b 23.5a 27.36a 27.16a 20.30de 20.60d 22.03c 21.76c

‘Nemark 49’ 11.56i 12.13i 14.63h 14.30h 16.53g 17.23f 20.6d 21.0e 23.9b 23.5a 26.96a 26.53a 19.96e 20.43de 21.66c 21.06cd Means with similar letters in each column are not significantly different by LSD multiple range test (p< 0.05)

Tab. 3. Comparison of interaction between variety, row spacing and different stages of maturity for B-carotene

Different stages green pink red deep red

Planting distances 25 33 41 49 25 33 41 49 25 33 41 49 25 33 41 49

Cultivar

‘Super A’ 51.0f 51.4f 57.4e 49.4f 258d 263c 288a 271.4b 24g 21.4h 7.4j 11i 0.50k 0.76k 0.50k 0.16k

‘1449’ 63.7g 66.7f 69.4e 64.4g 228d 238.7c 251.7b 264.7a 35.4g 27h 19.4i 17j 8.26k 9.7k 5.3l 7.4k

‘Nemark 49’ 36.4f 36f 35.7f 37.4f 313d 324c 342.4b 351a 58e 39e 31.7h 23.7i 11.4j 12.5j 7.6k 6.8k Means with similar letters in each column are not significantly different by LSD multiple range test (p< 0.05)

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fect of electrical conductivity fruit pruning and truss posi- tion on quality in green house tomato fruit. J Hortic Sci and Biotechnol 62:488-494.

Fernando C, Alisdair RF (2006). Metabolic regulation underly- ing tomato fruit development. J Experim Bot 57(9):1883- 1897.

Gaye MM, Jolliffe PA, Maurer AR (1992). Row cover and population density effects on yield of bell pepper in South Coastal British Columbia. Canadian J Pl Sci 72:901-909.

Gilmore AM, Yamamoto HY (1991). Resolution of lutein and zeaxanthin using a non-endcapped, lightly carbon-loaded c-18 high-performance liquid- chromatographic column. J Chromatogr 543:137-145.

Goodwin TW (1955). In: Paech K, Tracey MV (Eds.). Modern methods of plant analysis, Springer, Heidelberg 3:272.

Gould AA (1983). Tomato Production and quality evaluation.

AVI, New York, U.S.A, West port, Connecticut: AVI pub- lishing 478 p.

Huerres Perez C, Caraballo LN, Margolles J (1987). Effect of planting distance and number of plants per cluster in 3 cul- tivars of processing tomatoes (Lycopersicon esculentum Mill).

Centro Agricola 14:(3):21-30.

Konings E, Roomans H (1997). Evaluation and validation of an LC method for the analysis of carotenoids in vegetables and fruit. J Food Chem 59:599-603.

Laval-Martin D, Quennemet J, Moneger R (1975). Pigment evolution in Lycopersicon esculentum fruits during growth and development. Photochemistry 14:2357-2362.

Law-Ogbomo KE, Egharevba RKA (20090. Effects of planting density and NPK fertilizer application on yield and yield components of tomato (Lycospersicon esculentum Mill) in forest location. J Agric Sci 5(2):152-158.

McCoBum JP (1954). Effects of light on the formation of caro- tenoids in tomato fruits. Food Res 19:182.

Mahendran S, Bandara DC (2000). Effects of soil moisture stress at different growth stages on vitamin C, capsaicin, and β-carotene contents of chilli (Capsicum annum L.) fruits and their impact on yield. Trop Agric Res 12:95-106.

Malewski W, Markakfs P (1971). Ascorbic acid content of de- velop in tomato fruit. J Food Sci 36(3):537.

Manchanda AK, Bhopal S (1988). Effect of plant density and nitrogen on growth and fruit yield of bell pepper (Capsicum annuum L.). Indian J Agron 33(4):445-447.

Mauz F (1966). Flavor of fresh market tomato (Lycopersicon es- culentum Miil) as influence by harvest maturity and storage temperature. Ph. D thesis.

the amount of lycopene is influenced by the cultivar. An- derson (1969), Uddin et al. (1997) and Muhammad and Singh (2007) reported that plant spacing had a large affect on the PAR (photo synthetically active radiation) so the ratios of blue, far-red, red far and blue red as a result of decreased plant density were increased. This issue has an impact major on lycopen level. Also, Law and Eghrevba (2009) showed that tomato fruit exposed to sunlight dur- ing ripening contained more lycopene than those ripened in shade.

Conclusions

Several factors including: cultivar, row spacing and different stages of maturity can influence ascorbic acid, soluble solids, β-carotene and lycopene in tomato fruits.

According to this study, sunlight is a crucial factor on fruit quality. In general, increasing row spacing causes more light enter to the plant, so it increases ascorbic acid, lyco- pen and carotenoids content. In high row spacing, more photosynthesis take place because there is more light pen- etration in to the canopy and thus increases the amount of total soluble solids.

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Tab. 4. Comparison of interaction between variety, row spacing and different stages of maturity for lycopene

Different stages green pink red develop red

Planting distances 25 33 41 49 25 33 41 49 25 33 41 49 25 33 41 49

Cultivar

‘Super A’ 50k 55j 60i 68h 217g 223f 226e 235d 37c 371.7c 374.7b 377b 411.7a 411a 411.4a 409.7a

‘1449’ 32o 37.4n 42m 51.4l 165k 172g 183i 196.7h 301.7g 307.4f 311.4e 316.4d 361b 368.7a 357.4c 370.4a

‘Nemark 49’ 61.4m 73.4l 83k 98j 23i 246.4h 260.4g 277f 381e 389d 386.4d 386.4c 420.4b 431.4a 434.4a 418b Means with similar letters in each column are not significantly different by LSD multiple range test (p< 0.05)

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Tanaka M, Komochi S (1982). Research bulletin of Hokkaido National Agricultural Experimental Station 35:83-99.

Tonucci LH, Holden JM, Beecher GR, Khachik F, Davis CS, Mulokozi G (1995). Carotenoid content of thermally pro- cessed tomato-based food-products. J Agric Food Chem 43:579-586.

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Young TE, Juvik JA, Sullivan JG (1993). Accumulation of the components of total solids in ripening fruits of tomato. J Hort Sci 118(2):286-292.

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Spices. Tropical Agriculture Series, Vol. 1, Longman Group Ltd., London.

Shiraghinge FH, Mello PCT, Jacomino AP, Melo AMT, Purqu- erio LFV, Roquejani MS (2010). Yield and qualitative char- acterization of fresh market tomato hybrids of Italian and Santa Cruz type. J Hortic Bras 80:292-298.

Singh GH, Dillon TS (1986). Some aspects of seed extraction in tomato seed. Research 13(2):67-72.

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