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Effect of different fertilizers on growth and bioconstituents of red beet bulbs during three maturity stages.

INTRODUCTION

Red beet root (Beta vulgaris L. ssp. vulgaris, Chenopodiaceae) is a very important vegetable crop which exists all over the world. It ranks among the 10 most powerful vegetables due to its antioxidant capacity ascribed to the high polyphenolic and betalain contents. Total content of phenolics included a significant amount of phenolic acids viz p-coumaric, ferulic, p-hydroxybenzoic, protocatechuic, vanillic and syringic acids [1]. Betalains constituted red-violet betacyanins and yellow-orange betaxanthins. Especially betacyanins are the object of increased interest, because of their use for production of food colorants on a commercial scale. Additionally, previous studies reported a superior stability of betacyanins in comparison to betaxanthins [2].In addition, red beet bulbs were reported to contain alkaloids, saponins, flavonoids, tannins, vitamins and minerals [3, 4] and thus explaining their health promoting and prophylactic activities against numerous diseases from heart disease to cancer [5-6].

The problem associated with the use of synthetic chemical fertilizers is becoming a global one and researchers are working all over the world to find a solution to this problem. The excessive use of chemical fertilizer in agriculture causes environmental problems including soil, physical destruction and nutrient imbalance. As a consequence of the continually increasing demand for environmental protection and production of healthy food, it is necessary to increase the use of eco-friendly and environmentally safe natural fertilizers viz. organic, biofertilizers and yeast fertilizers. The main advantage of these natural fertilizers is that they do not pollute the soil and also do not show any negative effect to environment and human health. The vegetable fertilization must not corrupt the requirements of ecology and leads to increased plant productivity. Farmers and organizations must look forward for the suitable and optimal fertilization methods and fertilizer types. Finally obtaining fewer amounts of healthy products with less environmental disturbances should be preferred over obtaining higher amount of non-healthy products with more environmental disturbances.

Organic fertilizers are obtained from animal sources such as animal manure or plant sources like green manure. Continuous usage of inorganic fertilizers affects soil structure. Hence, organic manures can serve as alternative to mineral fertilizers for improving soil structure [7] and microbial biomass [8]. The addition of organic fertilizers to agricultural soils has beneficial effects on crop development and yields by improving soil physical and biological properties [9]. The suitability and usefulness of organic manure has been attributed to high availability of NPK content [10]. Organic and biofertilizers in comparison to the chemical fertilizers have lower nutrient content and slow release but they are as effective as chemical fertilizers over long periods of usage [11]. This was reported by some authors [12] who showed that organic fertilizers enhanced vegetative growth parameters and essential oil productivity of Pelargonium graveolens. Also, several researchers revealed that organic manuring increased the vegetative growth and biomass production effectively [13, 14].

Yeast and biofertilizers are live formulations of microorganisms obtained using natural election of different types of beneficial bacteria and fungi [15] that are ready to be used and improve the quality of the soil and the plant by increasing the nutrient availability. They naturally activate the microorganisms found in the soil restoring the soil's natural fertility and protecting it against drought and soil diseases and therefore stimulate plant growth [16]. Some authors [17] mentioned that yeast is an enriched source of phytohormones especially cytokinins, vitamins, enzymes, amino acids and minerals as well as that it has a stimulatory effect on the cell division and enlargement, protein and nucleic acids synthesis, chlorophyll formation and protective role against different stresses. Biofertilizer is a natural product carrying living microorganisms derived from the root or cultivated soil. So they don't have any ill effect on soil health and environment. Besides their role in atmospheric nitrogen fixation and phosphorous, these also help in stimulating the plant growth hormones providing better nutrient uptake and increased tolerance towards moisture stress. A small dose of biofertilizer is sufficient to produce desirable results because each gram of carrier of biofertilizers contains at least 10 million viable cells of a specific strain [18]. In our present investigation, a mixture of Azotobacter chroococcum, Bacillus megaterium and Bacillus circulans was used as biofertilizer where they secrete some organic acids which can solubilize P from insoluble and fixed forms to plant available forms and convert atmospheric N2 into plant available form of N in the soil. [19] These fertilizers are important factors in reducing the rate of using the chemical fertilizers to ensure safety for environment, improve soil fertility and increase soil productivity [20]. Additionaly, minimizing the consumption of chemical fertilizers was one of the targets of this work.

The aim of this work was to determine the effect of supplementing different fertilizers (organic, chemical, biofertilizer and yeast) and different harvesting stages on growth, bioconstituents (polyphenols and betanin) and in vitro antioxidant activity of red beet roots.

MATERIAL AND METHODS

Experimental site and plot layout:

Two field experiments were carried out at the Experimental Farm, Applied Research Center for Medicinal Plants, National Organization for drug Control and Research (NODCAR), Egypt, during 2011/2012 and 2012/2013 seasons to study the effect of some fertilizer treatments (organic, chemical, bio-fertilizers and yeast) on bulb growth and some chemical constituents of red beet bulbs. The seeds were planted using a randomized complete block design with three replications. Seeds were sown in sand loam soils on 15th October in both seasons in plots (1x1 m) containing four rows (25 cm in between) every row contains five hills (20 cm apart) and 45 days later, the plants were thinned, leaving only one seedling/hill. Physical and chemical characters of the used soil are shown in Tables (1) and (2), physical analysis was estimated according to [21] whereas, chemical analysis was determined according to [22].

Plant material:

Seeds of red beets (Beta vulgaris L. ssp. vulgaris) were obtained from Floriculture Farm, Horticulture Department, Faculty of Agriculture, Benha University and sown into the soil to produce a fully grown plant which was authenticated by Dr. Abd El Halim Abd El Mogali Mohamed, senior researcher, Flora and phytotaxonomy researches department, Horticultural research institute, Agricultural Research Center, Egypt.

Organic fertilizer treatment:

Organic fertilizer i.e. compost containing plant sources and cattle manure at the rate of 5 [m.sup.3]/fed., was thoroughly mixed with the soil before planting, the chemical properties of the tested compost are presented in Table (3). This treatment represented the control treatment in the present study.

Chemical fertilizer treatment: (NPK):

The plants were fertilized with full chemical fertilizer dose as a recommended dose; where ammonium nitrate (33.5% N) was added at the rate of 50Kg/fed., calcium superphosphate (15.5 % [P.sub.2][O.sub.5]) was added at the rate of 150 Kg/fed. and potassium sulphate (48.5 % [K.sub.2]O) at the rate of 100 Kg/fed. The amount of N and K fertilizers were divided into four equal portions as side dressing at 40, 50, 60 and 70 days after sowing date of both seasons. However, the amount of P-fertilizer was added to the soil before seed sowing during soil preparation.

Biofertilizer:

Red beet seeds were inoculated with a mixture of nitrobein, phosphorein and potassiumage that contained efficient strains of nitrogen fixing bacteria namely; Azotobacter chroococcum (1 x [10.sup.9] cfu/ml), phosphate dissolving bacteria: Bacillus megaterium var phosphaticum (1 x [10.sup.11] cfu/ml) and Bacillus circulans (1 x [10.sup.8] cfu/ml), respectively. The biofertilizer was supplied by the Department of Microbiology, faculty of Agriculture, Ain Shams University, Cairo. Seeds of red beet were washed with water and air-dried, thereafter they were soaked in cell suspension of the prementioned mixture for 30 min. Gum arabic (16 %) was added as an adhesive agent prior to soaking. The inoculated seeds were air dried at room temperature for one hour before sowing. Another two applications were applied at 4 (100%), 3(75%), 2(50%) and 1(25%) kg/fed as an aqueous solution, the first one was applied just before irrigation after 50 days from sowing date, whereas the second one was done after 70 days from sowing date to increase the power ability of bacteria.

Yeast fertilizer:

Yeast cultures were prepared by growing the yeast strain (Saccharomyces cervisiae) in yeast extract malt extract broth (YMB) (yeast extract 3 g/l, malt extract 3 g/l, peptone 5 g/l and glucose, 10 g/L) at 25 [+ or -] 1[degrees]C with shaking (150 rpm) for 48 to 72 h. The yeast cells were pelletized by centrifugation (5000 r.p.m) for 10 min and resuspended in sterilized tap water to the desired concentration (~[10.sup.8] cfu/ml).

In each season, yeast extract at a concentration of 4 (100%), 3 (75%), 2(50%) and 1(25%) l/fed., was added three times i.e., 30, 40 and 50 days after sowing date.

The treatments used in this work were combinations of the previously mentioned fertilizers i.e. chemical, bioferilizer and yeast, these treatments are listed in table (4).

Harvesting dates:

The plants were harvested at three different maturity stages i.e., the first one was after 2 months from sowing date(first stage), the second one was after 3 months from sowing date(second stage) whereas the third one was after 4 months from sowing date(third stage) in both seasons.

Determination of vegetative growth parameters:

Regarding the determination of vegetative growth parameters, at three stages, three plants from each plot were picked up randomly as representative samples for measuring and calculating the following characteristics: fresh weight (g), length (cm) and circumference (cm).

Chemicals and solvents:

Folin-Ciocalteu reagent; 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) (Sigma chemicals, USA); methanol; and HCl. (Labscan analytical sciences, Poland).

Reference authentic material:

Gallic acid (Sigma chemicals, USA) for determination of total polyphenolic activity.

Determination of betanin content:

For the betanin measurement, approximately 20 g of peeled and chopped bulbs were collected separately from each treatment for each replication and homogenized with a mortar and pestle. Then extracted with about 90 ml of 99% acidified water (with 1% HCl) by sonication for 10 minutes. The samples were filtered and the volume was adjusted to 100 ml with acidified water (stock solution). One ml of the filtrate was diluted to 10 ml with the same solvent. Betanin content was quantified by spectrophotometer (Unicam, UK) at 538 nm using absorptivity value (A1%) which was 1120, according to [23] and then converted into mg/100g FW.

Determination of total polyphenolic content:

Total polyphenols were determined by the method of [24] with some modifications and expressed as mg of gallic acid equivalent per 100g fresh matter. The total polyphenols content was estimated using Folin-Ciocalteu assay. The Folin-Ciocalteu reagent was added to a mixture of 0.25 ml sample (from a stock solution prepared in determination of betanin) and 1 ml of a sodium carbonate solution (4%). The volume was adjusted to 5 ml by distilled water. After 30 minutes, the absorbance was measured at 765 nm of wavelength against blank (composed of all reagents without the sample). The concentration of polyphenols was calculated from a standard curve plotted with known concentration of gallic acid (2-12 pg/ml).

Determination of in vitro antioxidant activity:

The antioxidant activity was evaluated adopting the radical scavenging DPPH method of [25] with slight modifications. Twenty grams of peeled and chopped bulbs were collected separately from each treatment for each replication and homogenized with a mortar and pestle. Then extracted with about 90 ml of methanol by sonication for 10 minutes. The samples were filtered and the volume was adjusted to 100 ml with methanol. Aliquots (0.05, 0.1 & 0.15 ml) of each extract were separately mixed with 5ml of a freshly prepared methanolic solution of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH, 20mg/l), completed to 10ml by methanol and left to stand for 30 minutes in the dark. The absorbance of samples was determined using a spectrophotomer at 517 nm and a control solution containing all reagents except the test sample was prepared. The radical-scavenging activity was calculated as a percentage of DPPH discoloration (I%) using the following equation:

I(%) = ([Abs.sub.control] - [Abs.sub.sample]/[Abs.sub.control]) x 100

Where: [Abs.sub.sampie] = absorbance of tested extracts,

[Abs.sub.Controi] = absorbance of control solution.

The [IC.sub.50], which is the concentration that causes 50% inhibition, was calculated for each case from the curve drawn for the I% of each sample versus the corresponding concentrations.

Statistical analysis:

All results are expressed as mean values [+ or -] standard error (SE). The obtained data (two seasons) were statistically analysed by one-way analysis of variance (ANOVA), using SPSS software version 20, followed by Dunette's test when comparing to control.

Results:

Effect of different fertilizers on bulb vegetative growth values:

Bulb fresh weight (g):

Data in table (4) showed that almost all treatments significantly increased bulbs weight of red beet compared to a control (treatment number 1) in both seasons. However, the heaviest bulbs fresh weight was scored by 100% chemical fertilizer (treatment number 2), followed by 75% biofertilization (treatment number 4) then the treatment of 75% yeast (treatment number 8). These treatments recorded the highest increments in this concern in the two seasons. Moreover the heaviest bulbs fresh weight was gained at the third stage harvest. Followed by the second one, whereas the lowest fresh weight was scored at the first stage harvest in the two seasons. Furthermore, all interactions between the studied fertilizers and harvest time significantly affected the bulb fresh weight of red beet, especially the interaction of 100% chemical fertilizer. Anyhow, the heaviest fresh weight of bulbs was registered by 100% chemical fertilizer at the third harvest in the seasons in the two seasons.

Bulb length (cm):

From table (4), the results revealed that bulb length of red beet was greatly influenced by the different treatments, especially the treatments number 2, 4, and 8 of 100% chemical fertilizer, 75% biofertilizer and 75% yeast, respectively, as compared with the control plants in the two seasons. Furthermore, the bulbs length increased with the advancement of harvesting date till reaching the maximum increase at the third stage in the two seasons. Moreover, almost all combinations between the studied fertilizers and harvest date increased the bulbs length, particularly those received 100% chemical fertilization and harvested after 4 months from planting in the two seasons, followed in descending order by those received 75% biofertilizer, then 75% yeast in the two seasons.

Bulb circumference (cm):

Data in table (4) reported that all applications significantly increased the circumference of red beet bulbs, particularly those received 100% chemical fertilizer, followed in descending order by the treatments number 4 and 8, of 75% biofertilizer and 75% yeast, respectively, in the two seasons. Moreover, there was a positive relationship between the values of bulb circumference and harvest date, while as the harvest date prolonged, the values of bulbs circumference increased to reach its maximum values at the third stage harvest in the two seasons. However, the highest values of bulbs circumference were scored by those received 100% chemical fertilizer, 75% biofertilizer and 75% yeast at the third stage harvest in both seasons.

Effect of different fertilizers on bulb bioconstituents:.

Betanin content:

From results in table (5), it was observed that almost all results of betanin content were significantly different than the control. The betanin content in all treatments was highest in the first stage then decreased in the second and the third stages, in both seasons. The betanin content significantly reached its maximum content in treatments number 4 among different concentrations of biofertilizers and 8 between the different yeast fertilizer treatments and even higher than the treatment with chemical fertilizer (number 2), when compared to the control treatment (number 1).

Total polyphenolic content:

Significant pronounced increase in total polyphenolic contents, in the red beet bulbs of all treatments, were obtained in both seasons when compared to control treatment. Results in table (5) revealed that , the highest significant polyphenolic contents were that produced by treatment number 4 among treatments of biofertilizers and treatment number 8 of the different yeast fertilizers. It was also observed that the polyphenolic content in the studied treatments increased in the bulbs of the second stage than those of the first stage then decreased in the third stage. Thus, the highest polyphenolic values of all treatments were obtained in the second stage in both seasons.

Antioxidant activity:

Data in table (5) showed that highest antioxidant activities in all treatments were exerted at the second stage in both seasons. The highest scavenging activities were afforded by treatments number 4 between the biofertilizer treatments and 8 among the yeast treatments. Similar to the results of the polyphenolic content, the antioxidant activities of the studied treatments increased in the second stage followed by a decrease in the activities in the third stage.

Discussion:

Red beet is a vegetable crop cultivated throughout the world as a source of food and natural dye. Medicinally, it has been used to treat a wide variety of ailments [5, 6]. The use of the suitable fertilizer in its cultivation and selecting the stage at which it should be harvested was the main objective in this study. Due to the increased attention which is now being paid to develop Integrated Plant Nutrition system (IPNS) that aims to enhance crop and soil productivity through a balanced use of all sources of nutrients, including chemical, organic and biofertilizers. The basic concept underlying the IPNS is the adjustment of soil fertility and plant nutrient supply to an optimum level for sustaining desired crop productivity through optimization of the benefits from all possible sources of plant nutrients in an integrated manner. We conducted a field experiment to evaluate the effects of the combined use of different concentrations of biofertilizer/yeast fertilizer along with chemical fertilizer. The results revealed that the treatment contained 75% biofertilizer and 25% chemical fertilizer showed significantly high vegetative growth parameters (weight, length and circumference of bulbs) compared to a control treatment and at the same time produced nearly similar results as that of the full chemical fertilizer. These results were higher than those obtained by full dose biofertilizer and that is in accordance with [26] who reported that biofertilizers cannot replace chemical fertilizers, but they can be used as a supplement along with chemical fertilizers. Also these results are in harmony with [27] who showed there was a 25% increase of lettuce yield for the treatment of A dose chemical fertilizer and biofertilizer compared to that of the full dose chemical fertilizer treatment, indicating that at least 50% of chemical fertilizer can be saved as multifunctional biofertilizer was used along with chemical fertilizer. Other authors [28] showed that combination use of biofertilizers and chemical fertilizers has increased fruit yield, individual fruit weight and total fruit weight of cucumber, while some authors [29] have indicated that the use of biofertilizer combined with chemical fertilizers has increased the shoot fresh weight and shoot dry weight of corn. Similarly, the combined treatment of 75% yeast and 25% chemical fertilizer significantly produced highest growth parameters amongst the different yeast treatments used in this study but yet less than the results of treatments number 4(75% biofertilizer and 25% chemical fertilizer) and 2 (full dose chemical fertilizer). These results are in disagreement with the results of the authors [30] who reported that plants fertilized with NPK alone gave the lowest records of all estimated pea yield parameters, whereas, plants fertilized with NPK in combination with yeast extract and micronutrients gave higher pods weight. This could be explained due to the presence of organic manure supplied to the field soil, which exaggerated the crop productivity with chemical fertilizer over the combination of yeast treatments and the chemical fertilizer. This explanation is in accordance with the authors [31] who reported that integrated supply of plant nutrients through farmyard manure and NPK fertilizer, along with Sesbania green manuring, played a significant role in sustaining soil fertility and crop productivity.

All studied treatments played an important role in determining and increasing the levels of betanin and polyphenolic contents of red beet bulbs. The values of the combined treatments of 75% biofertilizer/yeast and 25% chemical fertilizer were significantly higher as compared with control treatment.

These results are in line with those of [32] who stated that biofertilizers played a major role in determining the levels of phenolics, flavonoids and isoflavonoids of soybean seeds. The results also are in agreement with [33] who concluded that treating caraway plant by applying NPK (150 kg/ fed) combination with dry yeast (4 g/l) gave the best results regarding vegetative growth, seed yield, chemical composition, seed essential oil% and volatile oil constituents of caraway.

The antioxidant activity was high in all treatments as compared to control. The highest activities were produced by bulbs grown under combined treatments of 75% biofertilizer/yeast and 25% chemical fertilizer. These two treatments accelerated antioxidant activity greatly as compared to control. These high levels of antioxidant activity were associated with high values of betanin and polyphenols, which were known for their pronounced antioxidant activity [34, 35].

The highest vegetative growth parameters were observed in all treatments in the third stage, while maximum values of betanin were produced in the first stage and the highest results of polyphenols and antioxidant activities were attained in the second stage. Therefore, the present study strongly admit the harvest of the bulbs in the second stage in which somewhat high vegetative growth parameters were achieved together with considerable high amounts of bioconstituents. Also the combined treatments which could be preferably used in the cultivation are those containing 75% biofertilizer/yeast and 25% chemical fertilizer. Consequently, integrated application of chemical and biofertilizer/yeast due to their production of maximum growth parameters and bioactive metabolites could reduce the usage of a full dose chemical fertilizer and that may benefit human health.

Conclusion:

In conclusion, application of combination treatments of biofertilizers / yeast with chemical fertilizers as substitutes for full dose of inorganic fertilizers is recommended in order to grow the red beet bulbs. In addition application of similar treatments for cultivation of other medicinal and aromatic plants, should be considered as a main target in our life and a long term benefit to improve environmental conditions and human health. Meanwhile, red beet bulbs should be harvested at the appropriate level of maturity, considering growth and bioconstituents, in order to achieve considerably high contents of biologically active constituents and lower production cost.

Received 28 December 2015; Accepted 28 January 2016; Available online 24 February 2016

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Ghada Farouk and Horia Sharawy

Department of medicinal plants and natural products, National Organization for Drug Control and Research (NODCAR), Giza, Egypt. Address For Correspondence:

Ghada Farouk, Department of medicinal plants and natural products, National Organization for Drug Control and Research (NODCAR), Giza, Egypt.
Table 1: Physical analysis of the experimental soil:

Parameters       Results

Coarse sand(%)   18.07
Fine sand(%)     69.5
Silt(%)          3.1
Clay(%)          8.7
Textural class   Loamy sand

Table 2: Chemical analysis of the experimental soil.

Soluble cations     Soluble anions            Macroelements
(m.equ/L)           (m.equ/L)                 (ppm)

[Ca.sup.++]   1.4   HC[O.sup.-.sub.3]   0.8   Total N              10
[Mg.sup.++]   0.8   [Cl.sup.-]          1.5   [P.sub.2][O.sub.5]   5
[Na.sup.+]    1.8   S[O.sup.2-.sub.4]   3.1   [K.sub.2]O           388
[K.sup.+]     1.4

Microelements       pH    EC            CaC[O.sub.3]
(ppm)                     (m.mohs/cm)

Fe            3     7.8   0.55          3.2
Cu            0.25
Zn            0.98
Mn            4.4

Table 3: Chemical properties
of the used compost:

Parameters           Results

Ec (dS.[m.sup.-1])   2.94
pH                   6.73
Total C (%)          23.14
Total N (%)          1.28
Total P (%)          0.69
Total K (%)          1.59
Total Fe (ppm)       1425
Total Zn (ppm)       384
C:N ratio            18:1

Table 4: Treatments used in this experiment

Number of     Chemical     Biofertilizer   Yeast
treatment     fertilizer   (%)             fertilizer
              (%)                          (%)

1 (Control)   --
2             100          --              --
3             --           100             --
4             25           75              --
5             50           50              --
6             75           25              --
7             --           --              100
8             25           --              75
9             50           --              50
10            75           --              25

Table 4: Effect of different fertilizers on bulb weight,
length and circumference

No. of      Bulb weight(g)
treatment

            First season

            1st stage                2nd stage

1           23.83 [+ or -] 0.71      98.97 [+ or -] 1.30
2           130.37 [+ or -] 2.86 *   298.73 [+ or -] 1.79 *
3           41.03 [+ or -] 2.23 *    187.40 [+ or -] 1.00 *
4           102.57 [+ or -] 2.15 *   295.60 [+ or -] 3.92 *
5           64.23 [+ or -] 1.08 *    156.60 [+ or -] 11.68 *
6           63.57 [+ or -] 2.04 *    165.93 [+ or -] 2.70 *
7           40.10 [+ or -] 1.86 *    180.83 [+ or -] 1.53 *
8           77.37 [+ or -] 2.00 *    235.57 [+ or -] 1.80 *
9           75.10 [+ or -] 1.55 *    216.90 [+ or -] 3.27 *
10          48.07 [+ or -] 2.67 *    166.13 [+ or -] 1.31 *

No. of      Bulb weight(g)
treatment

            First season             Second season

            3rd stage                1st stage

1           140.30 [+ or -] 8.59     22.05 [+ or -] 2.12
2           491.77 [+ or -] 1.86 *   81.56 [+ or -] 2.96 *
3           345.83 [+ or -] 2.91 *   34.75 [+ or -] 1.59 *
4           455.63 [+ or -] 8.13 *   56.77 [+ or -] 1.93 *
5           309.10 [+ or -] 2.20 *   37.33 [+ or -] 0.82 *
6           193.03 [+ or -] 1.81 *   36.57 [+ or -] 1.67 *
7           342.10 [+ or -] 2.14 *   41.93 [+ or -] 2.00 *
8           423.20 [+ or -] 1.44 *   52.90 [+ or -] 2.25 *
9           269.50 [+ or -] 1.82     35.27 [+ or -] 2.10 *
10          286.67 [+ or -] 1.58 *   46.80 [+ or -] 1.68 *

No. of      Bulb weight(g)
treatment

            Second season

            2nd stage                3rd stage

1           170.27 [+ or -] 3.05     343.40 [+ or -] 1.33
2           385.83 [+ or -] 5.32 *   745.73 [+ or -] 10.47 *
3           193.60 [+ or -] 5.31 *   539.10 [+ or -] 4.58 *
4           232.5 [+ or -] 1.46 *    646.50 [+ or -] 2.37 *
5           178.63 [+ or -] 4.09     609.60 [+ or -] 5.77 *
6           168.67 [+ or -] 0.77     528.00 [+ or -] 1.21 *
7           220.20 [+ or -] 3.29 *   601.07 [+ or -] 2.70 *
8           224.67 [+ or -] 3.93 *   628.87 [+ or -] 1.86 *
9           171.33 [+ or -] 2.09     372.70 [+ or -] 1.65 *
10          214.33 [+ or -] 1.99 *   595.73 [+ or -] 2.98 *

No. of      Bulb length (cm)
treatment

            First season

            1st stage               2nd stage

1           11.33 [+ or -] 0.33     12.67 [+ or -] 1.45
2           16.33 [+ or -] 1.26 *   19.67 [+ or -] 0.48 *
3           13.00 [+ or -] 0.29     18.00 [+ or -] 0.58 *
4           15.67 [+ or -] 0.73 *   19.33 [+ or -] 0.45 *
5           13.33 [+ or -] 0.44     16.00 [+ or -] 0.58
6           11.67 [+ or -] 0.88     15.67 [+ or -] 0.67
7           12.83 [+ or -] 0.39     18.50 [+ or -] 0.87 *
8           14.50 [+ or -] 0.76 *   18.67 [+ or -] 0.19 *
9           14.17 [+ or -] 0.77     16.50 [+ or -] 0.29 *
10          13.50 [+ or -] 0.75     15.67 [+ or -] 1.20

No. of      Bulb length (cm)
treatment

            First season            Second season

            3rd stage               1st stage

1           14.00 [+ or -] 0.58     7.83 [+ or -] 0.73
2           23.67 [+ or -] 0.99 *   11.83 [+ or -] 0.93 *
3           19.00 [+ or -] 0.58 *   10.17 [+ or -] 0.17
4           22.50 [+ or -] 1.10 *   11.67 [+ or -] 0.88 *
5           18.00 [+ or -] 0.58 *   9.50 [+ or -] 0.50
6           17.33 [+ or -] 0.88 *   8.50 [+ or -] 0.29
7           20.00 [+ or -] 0.58 *   11.33 [+ or -] 0.88 *
8           21.00 [+ or -] 0.58 *   11.50 [+ or -] 0.29 *
9           18.50 [+ or -] 0.87 *   10.17 [+ or -] 0.60
10          17.67 [+ or -] 0.52 *   9.50 [+ or -] 0.29

No. of      Bulb length (cm)
treatment

            Second season

            2nd stage               3rd stage

1           15.67 [+ or -] 0.67     19.33 [+ or -] 0.33
2           23.33 [+ or -] 1.01 *   25.67 [+ or -] 0.88 *
3           19.33 [+ or -] 0.44 *   22.00 [+ or -] 1.15
4           22.67 [+ or -] 0.72 *   24.67 [+ or -] 1.48 *
5           16.00 [+ or -] 0.58     23.00 [+ or -] 1.15
6           19.33 [+ or -] 0.33 *   24.00 [+ or -] 1.15 *
7           18.00 [+ or -] 1.50     20.33 [+ or -] 1.20
8           19.67 [+ or -] 1.30 *   24.50 [+ or -] 1.04 *
9           18.50 [+ or -] 0.87     23.33 [+ or -] 0.60
10          18.67 [+ or -] 0.88     21.67 [+ or -] 1.20

No. of      Bulb circumference (cm)
treatment

            First season

            1st stage               2nd stage

1           11.33 [+ or -] 0.73     20.50 [+ or -] 0.87
2           18.23 [+ or -] 0.93 *   32.17 [+ or -] 1.74 *
3           13.00 [+ or -] 0.76     23.50 [+ or -] 1.26
4           17.23 [+ or -] 0.53 *   27.00 [+ or -] 1.15 *
5           16.17 [+ or -] 0.73 *   25.17 [+ or -] 1.59 *
6           15.73 [+ or -] 0.37 *   24.00 [+ or -] 0.76
7           13.03 [+ or -] 1.36     23.67 [+ or -] 0.73
8           16.50 [+ or -] 0.87 *   25.17 [+ or -] 1.01 *
9           17.17 [+ or -] 1.04 *   25.00 [+ or -] 0.76
10          11.93 [+ or -] 0.47     21.00 [+ or -] 0.57

No. of      Bulb circumference (cm)
treatment

            First season            Second season

            3rd stage               1st stage

1           21.00 [+ or -] 0.58     10.50 [+ or -] 0.29
2           32.17 [+ or -] 0.60 *   17.33 [+ or -] 1.01 *
3           23.83 [+ or -] 0.17     14.00 [+ or -] 1.15 *
4           30.50 [+ or -] 0.76 *   15.67 [+ or -] 0.33 *
5           26.50 [+ or -] 0.76 *   13.50 [+ or -] 0.76
6           28.17 [+ or -] 0.73 *   13.17 [+ or -] 0.60
7           28.17 [+ or -] 1.01 *   13.50 [+ or -] 0.87
8           28.50 [+ or -] 0.87 *   14.00 [+ or -] 0.58 *
9           24.25 [+ or -] 1.09     12.50 [+ or -] 0.29
10          27.25 [+ or -] 1.01 *   13.67 [+ or -] 0.88 *

No. of      Bulb circumference (cm)
treatment

            Second season

            2nd stage               3rd stage

1           21.83 [+ or -] 0.29     26.67 [+ or -] 0.73
2           29.00 [+ or -] 1.01 *   36.83 [+ or -] 1.09 *
3           21.67 [+ or -] 1.15 *   33.33 [+ or -] 0.88 *
4           26.17 [+ or -] 0.33 *   36.83 [+ or -] 0.73 *
5           22.00 [+ or -] 0.76     34.67 [+ or -] 1.45 *
6           22.00 [+ or -] 0.60     32.33 [+ or -] 1.20 *
7           24.33 [+ or -] 0.87     30.83 [+ or -] 0.83
8           24.83 [+ or -] 0.58 *   35.17 [+ or -] 0.44 *
9           22.33 [+ or -] 0.29     34.33 [+ or -] 1.76 *
10          24.33 [+ or -] 0.33     33.17 [+ or -] 1.36 *

Values are mean of three results [+ or -] SE; * Statistically
significant from control at P<0.05

Table 5: Effect of different fertilizers on betanin,
polyphenolic content and antioxidant activity of red
beet bulbs

No. of      Betanin (mg/100g F.wt.)
treatment

            First season

            1st stage                2nd stage

1           43.19 [+ or -] 1.44      29.51 [+ or -] 1.45
2           85.41 [+ or -] 1.56 *    72.38 [+ or -] 1.39 *
3           85.83 [+ or -] 1.70 *    42.05 [+ or -] 0.40 *
4           87.49 [+ or -] 0.97 *    45.92 [+ or -] 1.82 *
5           77.93 [+ or -] 1.75 *    42.95 [+ or -] 0.75 *
6           74.75 [+ or -] 1.79 *    44.61 [+ or -] 0.08 *
7           85.41 [+ or -] 1.84 *    55.25 [+ or -] 1.50 *
8           105.19 [+ or -] 0.98 *   53.36 [+ or -] 0.23 *
9           87.97 [+ or -] 1.58 *    44.00 [+ or -] 1.38 *
10          76.14 [+ or -] 1.01 *    48.44 [+ or -] 1.98 *

No. of      Betanin (mg/100g F.wt.)
treatment

            First season            Second season

            3rd stage               1st stage

1           22.57 [+ or -] 0.46     34.78 [+ or -] 1.47
2           28.78 [+ or -] 1.90 *   56.79 [+ or -] 1.98 *
3           27.37 [+ or -] 0.22     78.90 [+ or -] 0.85 *
4           32.74 [+ or -] 2.00 *   106.55 [+ or -] 2.00 *
5           31.37 [+ or -] 0.92 *   69.75 [+ or -] 0.76 *
6           23.75 [+ or -] 1.67 *   87.72 [+ or -] 1.28 *
7           52.60 [+ or -] 0.59 *   62.13 [+ or -] 0.84 *
8           37.60 [+ or -] 0.06 *   76.83 [+ or -] 2.01 *
9           37.08 [+ or -] 1.41 *   73.68 [+ or -] 0.90 *
10          33.60 [+ or -] 0.41 *   67.80 [+ or -] 1.13 *

No. of      Betanin (mg/100g F.wt.)
treatment

            Second season

            2nd stage                3rd stage

1           24.66 [+ or -] 2.01      11.77 [+ or -] 1.95
2           40.34 [+ or -] 1.13 *    9.27 [+ or -] 1.04
3           32.17 [+ or -] 0.71 *    15.03 [+ or -] 1.79
4           43.78 [+ or -] 1.94 *    34.40 [+ or -] 1.16 *
5           39.93 [+ or -] 1.07 *    16.01 [+ or -] 0.81
6           59.81 [+ or -] 1.28 *    10.65 [+ or -] 1.81
7           38.33 [+ or -] 1.39 *    12. 41[+ or -] 1.0 0
8           52.05 [+ or -] 1.27 *    17.47 [+ or -] 0.98 *
9           38.33 [+ or -] 1.95 *    10. 39[+ or -] 0.5 7
10          34.18 [+ or -] 1.21 *    16. 18[+ or -] 2.6 3

No. of      Polyphenolic content(mg/100g F.wt.)
treatment

            First season

            1st stage                 2nd stage

1           65.17 [+ or -] 0.43       83.53 [+ or -] 1.32
2           84.40 [+ or -] 0.95 *     120.73 [+ or -] 1.60 *
3           77.47 [+ or -] 1.47 *     145.93 [+ or -] 0.79 *
4           93.90 [+ or -] 0.78 *     226.00 [+ or -] 0.64 *
5           79.27 [+ or -] 0.61 *     182.37 [+ or -] 0.52 *
6           83.77 [+ or -] 0.89 *     144.25 [+ or -] 1.65 *
7           98.60 [+ or -] 0.35 *     141.9 [+ or -] 1.33 *
8           113.37 [+ or -] 0.26 *    163.80 [+ or -] 1.04 *
9           104.80 [+ or -] 0.59 *    138.20 [+ or -] 0.46 *
10          104.03 [+ or -] 1.87 *    151.17 [+ or -] 1.29 *

No. of      Polyphenolic content (mg/100g F.wt.)
treatment

            First season              Second season

            3rd stage                 1st stage

1           71.60 [+ or -] 1.87       71.98 [+ or -] 1.00
2           66.87 [+ or -] 1.88       97.67 [+ or -] 0.82 *
3           94.03 [+ or -] 1.94 *     70.52 [+ or -] 1.88
4           170.3 0[+ or -] 1.10 *    106.25 [+ or -] 1.93 *
5           112.2 7[+ or -] 0.33 *    95.83 [+ or -] 0.97 *
6           122.5 [+ or -] 1.73 *     115.40 [+ or -] 1.06 *
7           97.50 [+ or -] 0.65 *     80.17 [+ or -] 0.19 *
8           108.97 [+ or -] 0.97 *    115.84 [+ or -] 1.03 *
9           43.21 [+ or -] 1.51 *     83.01 [+ or -] 0.73 *
10          113.80 [+ or -] 0.67 *    91.59 [+ or -] 1.20 *

No. of      Polyphenolic content (mg/100g F.wt.)
treatment

            Second season

            2nd stage                 3rd stage

1           102.30 [+ or -] 1.07      44.72 [+ or -] 0.26
2           117.85 [+ or -] 0.93 *    74.04 [+ or -] 0.48 *
3           188.08 [+ or -] 1.11 *    45.50 [+ or -] 0.37
4           140.48 [+ or -] 0.89 *    57.59 [+ or -] 0.54 *
5           169.91 [+ or -] 0.11 *    61.61 [+ or -] 0.77 *
6           137.30 [+ or -] 1.29 *    67.96 [+ or -] 0.42 *
7           80.78 [+ or -] 0.57 *     73.09 [+ or -] 2.11 *
8           139.92 [+ or -] 0.34 *    104.08 [+ or -] 0.29 *
9           99.34 [+ or -] 0.90       59.15 [+ or -] 0.55 *
10          117.01 [+ or -] 0.82 *    88.47 [+ or -] 0.06 *

No. of      Antioxidant activity (IC 50: mg/ml F.wt.)
treatment

            First season            Second season

            1st     2nd     3rd     1st     2nd     3rd
            stage   stage   stage   stage   stage   stage

1           3.83    1.97    3.07    3.00    2.75    8.9
2           2.34    2.09    3.53    9.53    4.96    9.1
3           2.67    1.78    2.28    10.4    5.18    13.73
4           1.38    0.67    2.00    2.77    1.92    7.13
5           2.30    2.03    2.59    6.08    5.11    20.38
6           3.10    1.77    2.85    8.55    3.07    30.98
7           2.12    2.10    4.04    2.88    2.27    7.41
8           1.87    0.39    2.31    1.78    1.26    6.29
9           3.19    1.25    6.24    4.14    3.03    8.55
10          2.49    1.89    5.80    14.53   2.00    11.01

Values are mean of three results [+ or -] SE except [IC.sub.50]
calculated from curves of I% vs concentrations; * Statistically
significant from control at P [less than or equal to] 0.05
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Author:Farouk, Ghada; Sharawy, Horia
Publication:Advances in Environmental Biology
Article Type:Report
Date:Jan 1, 2016
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