Oriental Journal of Chemistry

Vol. 26(4), 1353-1360 (2010)

The potential of seaweed liquid fertilizer on the growth and antioxidant enhancement of Helianthus annuus L.
N. AKILA and T. JEYADOSS*
Department of Chemistry and Biosciences, Srinivasa Ramanujan Centre, SASTRA University, Kumbakonam - 612 001 (India). (Received: September 03, 2010; Accepted: October 11, 2010) ABSTRACT
A field experiment was carried out during the chithiraipattam (April-May) in India in 2010 to study the effects of foliar applications of different concentrations of seaweed liquid fertilizer (SLF) of Sargassum wightii on growth, biochemical constituents and antioxidant enzymes of Helianthus annuus L. to explore the possibilities of reducing or avoiding chemical fertilizers and to obtain the highest growth and productive parameters. The foliar spray was given twice in 15days interval at two concentrations (2.5%, 5.0%) of SLF. The aqueous extract of Sargassum wightii was found to promote almost all the parameters such as shoot length, root length, leaf length, leaf breadth, fresh weight, dry weight and number of leaves. The biochemical contents such as chlorophyll a, b, total chlorophyll, carotenoids, carbohydrate, total phenols and antioxidant enzymes such as catalase, peroxidase and glutathione peroxidase (GPX) were also found to be higher in the leaves. Among the two concentrations 2.5% was found to produce better results in almost all the parameters. The results revealed that foliar application of seaweed extract could be a promising option for pollution free non toxic growth enhancement of Helianthus annuus L.

Key words: Sargassum wightii, growth, biochemical constituents, antioxidant enzymes, Seaweed Liquid Fertilizer.

INTRODUCTION Any improvement in agricultural system resulting in higher production should reduce the negative environmental impact on agriculture and enhance the sustainability of the system. One such approach is the use of biostimulants, which can successfully replace harmful and costly inorganic fertilizers. Marine bioactive substances extracted from marine algae are used in agricultural and horticultural crops (Blunden, 1991). Seaweeds are the macroscopic marine algae found at the bottom in relatively shallow coastal waters. They grow in shallow and deep sea areas up to 180 meter depth and in baers

on the solid substrate such as rocks, dead corals and pebbles. The seaweeds are totally different from higher plants as they neither have true leaves, stems and roots or vascular systems nor specialized sex organs. Seaweeds have been used as manure, cattle feed, food for human consumption and as a source of phycocolloids such as agar, alginic acid and carrageenan (Chapman, 1970). Besides their application as farmyard manure (FYM), liquid extracts obtained from seaweeds (LSF/SLF) have recently gained importance as foliar sprays for several crops (Metha et al., 1967; Bokil et al., 1974).

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Akila & Jeyadoss, Orient. J. Chem., Vol. 26(4), 1353-1360 (2010) Seeds Viable seeds were obtained from the Tamil Nadu Agricultural University, Coimbatore. Care was taken in selecting the seeds of uniform size and they were stored in a metal tin as suggested by Rao (1976). Experimental design and treatment The experiment compr ised of four treatments, viz, 0 (control, water spray), 2.5%, 5% (volume/volume) of seaweed extract in water and 0.1% NPK spray. Two sprays of Sargassum wightii derived extract and NPK were applied, one at the seedling stage (15th days after sowing) and the second at 15days after first spray (30th days after sowing). The treatments were distributed in a randomized block design with three replications. Soil analysis The soil sample used for field experiment was subjected to the analysis of physico-chemical properties. The parameters like coarse sand, texture class, nitrogen, phosphorus, potassium, EC, Ph, organic carbon, organic matter were found out. Morphological parameters The morphological characters such as shoot length, root length, leaf length, leaf breadth, fresh weight, dry weight, number of leaves were measured on 30th and 45th day after sowing. Biochemical estimation The biochemical constituents such as chlorophyll a, b, total chlorophyll (Arnon, 1949), carotenoid (Kirk and Allen, 1965), carbohydrate (Hedge and Hofreiter, 1962) and total phenols (Malic and Singh, 1980) were also determined. Antioxidant enzyme assay Catalase activity was estimated by the method of Barber (1980), peroxidase was determined by Rodney and Boyer (1996) and glutathione peroxidase were analyzed by Overbaugh and Fall (1982). Plants from each treatment were randomly drawn for various analysis. Plant from the field were uprooted carefully and washed in tap water. They were then processed for different analysis. All the growth parameters, biochemical constituent and

The application of seaweed fertilizer for different crops was of great importance to substitute the commercial chemical fertilizers and to reduce the cost of production. Liquid fertilizers derived from seaweeds are found to be superior to chemical fertilizers due to high level of organic matter, macro and micro nutrients, amino acids, vitamins, cytokinins, auxin and abscisic acid like growth promoting substances (Mooney and Van Staden, 1986). It was also used to develop tolerance to environment stress (Zhang et al ., 2003) and increase nutrient uptake from soil (Verkleij, 1992; Turan and Kose, 2004). The present study was undertaken to investigate the effect of seaweed liquid fer tilizers on the growth, biochemical constituents and antioxidant enzymes of Helianthus annuus L. MATERIAL AND METHODS Collection of Seaweeds The seaweed used in the present study was Sargassum wightii belonging to the class Phaeophyceae. They were collected from the coastal area of Rameswaram, India during March 2010. The algal species were hand picked and washed thoroughly with seawater to remove all the unwanted impurities, adhering sand particles and epiphytes. It was kept in an ice box containing slush ice and transpor ted to the laboratory. Samples were washed thoroughly using tap water to remove the salt on the surface of the sample. The water was drained off and the algae were spread on blotting paper to remove excess water. Preparation of seaweed liquid fertilizer One kg of seaweed was cut into small pieces and boiled separately with distilled water for an hour and filtered. The filtrate was taken as 100% concentration of the seaweed extract and from this different concentrations (2.5%, 5.0%) were prepared using distilled water (Bhosle et al., 1975). As the SLF contain organic matter, the seaweed liquid fertilizer were refrigerated between 0 and 40C. Test crop plant The crop plant, selected for the present study was Helianthus annuus L. belonging to the family Asteraceae.

Akila & Jeyadoss, Orient. J. Chem., Vol. 26(4), 1353-1360 (2010) antioxidant enzymes were analysed on 30th and 45th day after sowing. RESULTS A physico-chemical analysis of the experimental soil is presented in Table -1. The soil of the site was clay loam with pH 8.0. The experimental soil was found to contain 0.19% organic carbon, Nitrogen (60ppm), Phosphorous (11.1ppm), Potassium (90ppm) and organic matter 2.8%. Table 1: Physico - chemical analysis of the experimental soil Properties Physical analysis Coarse sand (%) Texture class Chemical analysis N (ppm) P (ppm) K (ppm ) EC (dsm-1) pH Organic carbon % Organic matter % Values

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sowing (DAS) is presented in Table-4. The activities of antioxidant enzymes such as catalase, peroxidase and glutathione peroxidase were found to be much higher in plant treated with 2.5% concentration when compared to control, 5% concentration of SLF and chemical fertilizer. DISCUSSION The present investigation has been under taken to achieve higher production of Helianthus annuus .L with eco-friendly and completely safe SLF in the place of costly, harmful and toxic chemical fertilizers. Bhosle e t a l . ( 1 9 7 5 ) p r e p a r e d a s e aw e e d l i q u i d fer tilizer and studied its effect on Phaeseolus vulgar is . Rama Rao (1991) repor ted good yields of Zizyphus rugosa fruits, where leaf spray of SLF obtained from Sargassum wightii was used . In the present investigation it is quite interesting to mention here that 2.5% c o n c e n t ra t i o n o f S L F h a s r e m a r k a bl e influence on all the growth parameters such as shoot length, root length, leaf length, leaf breadth, fresh weight, dr y weight and number of leaves which is in conformity with a earlier repor t of Metha et al . (1967). It is also wor th mentioning here that 5% SLF has also been found to have more or less equal growth promoting influence. Stephenson (1974) recorded that lower concentration of SLF prepared from Ascopyllum and Laminaria accelerated the growth in maize. These results observed by us are in agreement with those of Chandrasekar et al . (2005) who reported that the maximal plant height, leaf area and leaf length and number of leaves were observed in the plot treated with the biofertilizer. Rama Rao (1991) reported that the dry powder and liquid formulators of the seaweed sargassum wightii increase the fresh weight of tomato plants. The number of leaves and leaf area are important factors, because the leaves are the structures bearing photosynthetic machinery and an increased in leaf number and area, may promote

8.4 clay loam 60 11.1 90 0.14 8.0 0.19 2.8

The effect of SLF of Sargassum wightii on growth parameters such as shoot length, root length, leaf length, leaf breadth, fresh weight, dry weight and number of leaves recorded at 30th and 45th days after sowing (DAS) is presented in Table-2. The data of various biochemical constituents such as chlorophyll a, b, total chlorophyll, carotenoid, carbohydrate and total phenols observed on 30th and 45th days after sowing (DAS) are presented in Table-3. Even though both the concentrations enhanced the formation of all the biochemical constituents, comparatively a marked increase in all biochemical content have been observed in 2.5% concentration of SLF. The effect of SLF of Sargassum wightii on antioxidant enzymes of the leaves of Helianthus annuus.L measured on 30th and 45th days after

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Table 2: Effect of SLF of Sargassum wightii on growth parameters of Helianthus annuus.L 30DAS SLF (2.5%) 89.3±0.2 12.5±0.1 11.5±1.2 6.7±0.4 60.2±1.0 6.03±0.04 18±1.0 SLF (5.0%) Chemical fertilizer Control SLF (2.5%) SLF (5.0%) 45DAS Chemical fertilizer

Parameters

Control

Shoot length(cm)

45.3±0.2

Root length (cm)

8.8±0.1

Leaf length (cm)

9±1.0

Leaf breadth (cm)

4.7±0.2

Fresh weight (g)

30±1.0

Dry weight (hg

3.93±0.01

Akila & Jeyadoss, Orient. J. Chem., Vol. 26(4), 1353-1360 (2010)

No of leaves

10±2.0

56.2±0.3 (24.06) 9.8±0.1 (11.36) 15±1.0 (66.66) 9.1±0.2 (93.61) 58.5±1.5 (95.0) 6.82±0.01 (75.53) 17±1.0 (70.0)

55.3±0.3 (22.07) 8.5±0.1 (-3.40) 14.5±1.5 (61.11) 7.3±0.05 (24.06) 56.3±1.0 (86.66) 5.76±0.01 (46.56) 15±1.0 (50.0)

52±0.1 (14.79) 8.9±0.1 (1.13) 13.5±1.3 (50.0) 7.5±0.1 (24.06) 48±1.6 (60.0) 5.79±0.02 (47.32) 14±1.0 (40.0)

106±0.4 (18.70) 18±0.1 (44.0) 16.2±1.0 (40.86) 12.1±0.2 (80.59) 224±1.2 (272.09) 30±0.01 (397.51) 24±2.0 (33.33)

101.3±0.03 (13.43) 17.1±0.1 (36.8) 14.5±1.0 (26.08) 11.8±0.0 (76.11) 219±1.0 (263.78) 27±0.01 (347.76) 21±1. 0 (16.66)

99.1±0.1 (10.99) 17.8±0.1 (42.4) 13.4±1.3 (16.52) 12.0±0.3 (79.10) 208±1.6 (245.51) 24±0.02 (298.0) 22±0.03 (22.22)

- Values are expressed as mean ± SD DAS - days after sowing

SLF - Seaweed Liquid Fertilizer

Table 3: Effect of SLF of Sargassum wightii on biochemical composition of the leaves of Helianthus annuus.L 30DAS SLF (2.5%) 2.56±0.05 1.01±0.02 3.50±0.005 2.70±0.04 0.14±0.03 2.57±0.02 SLF (5.0%) Chemical fertilizer Control SLF (2.5%) SLF (5.0%) 45DAS Chemical fertilizer

Parameters

Control

1.65±0.005

0.65±0.02

2.33±0.01

2.72±0.05

0.45±0.03

Chlorophyll “a” (mg g-1FW) Chlorophyll “b” (mg g-1FW) Total Chlorophyll (mg g-1FW) Carotenoid (mg g-1FW) Carbohydrate (mg g-1 FW) Total Phenols (mg g-1FW)

1.06±0.10

2.07±0.005 (25.45 ) 0.74±0.01 (13.84 ) 2.73±0.03 (17.16) 3.12±0.05 (14.70 ) 0.61±0.01 (35.55 ) 1.94±0.03 (83.01 )

1.90±0.07 (15.15 ) 0.66±0.01 (1.53 ) 2.53±0.03 (8.58) 3.01±0.05 (10.66 ) 0.58±0.06 (28.88 ) 1.69±0.04 (59.43 )

1.92±0.01 (16.36 ) 0.64±0.05 (1.53 ) 2.48±0.01 (6.43) 3.04 ±0.01 (11.76 ) 0.56±0.03 (24.44 ) 1.72±0.06 (62.26 )

2.95±0.03 (15.23) 1.21±0.04 (19.80) 4.06±0.01 (16.0 ) 2.74±0.003 (1.48 ) 0.43±0.02 (207.14 ) 3.54±0.05 (37.74 )

2.90±0.06 (13.28) 1.19±0.01 (17.82 ) 3.99±0.02 (14.0 ) 2.72±0.005 (0.74 ) 0.41±0.03 (192.85 ) 3.44±0.03 (33.85 )

2.40±0.01 (6.25 ) 1.08±0.07 (6.93 ) 2.68±0.05 (23.42 ) 2.68±0.02 (0.74 ) 0.43±0.06 (207.14 ) 3.47±0.01 (35.01 )

Akila & Jeyadoss, Orient. J. Chem., Vol. 26(4), 1353-1360 (2010)

- Values are expressed as mean ± SD DAS - days after sowing

SLF - Seaweed Liquid Fertilizer

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Akila & Jeyadoss, Orient. J. Chem., Vol. 26(4), 1353-1360 (2010) the better translocation of water uptake and deposition of nutrients and yield (Jeevajyothi et al ., 1993). It is indeed worth mentioning that there is a marked improvement in all the above parameters with SLF fo l i a r s p r ay administration. The SLF had also influence in improving biochemical constituents such as chlorophyll a, b, total chlorophyll, carotenoids and total phenols content of experimental plants at lower concentration of 2.5%. The marked increase in the chlorophyll content observed in the present study has also been reported in the observations made by Whapman et al. (1993) and Blunden et al . (1997). Mostafa and Zheekh (1999) reported that the plant growth substance present in seaweed liquid fertilizer might have enhanced the chlorophyll content in the leaves. The seaweed extract applied as foliar spray enhanced the leaf chlorophyll level in plants (Blunden et al ., 1996). In the present investigation, plants treated with lower concentration (2.5%) of seaweed liquid fertilizers were found to show better response in terms of enzyme activities such as catalase, peroxidase and glutathione peroxidase. These results coincide with the studies on Vigna Zeamays (Linga Kumar et al., 2002 and Asir Selin Kumar et al ., 2007). Nor mally the higher activity of the enzyme appears to be the result of greater mobilization of storage compound and energy sources (Bardzik et al., 1971). I t i s c o n c l u d e d f r o m t h e a b ove discussion that in Helianthus annuus.L seaweed liquid fer tilizer stimulates significantly almost all parameters of growth, b i o c h e m i c a l c o n s t i t u e n t s a n d a n t i ox i d a n t activities in field conditions, when compared to control and chemical fer tilizer. This simple practice of application of eco-fr iendly seaweed liquid fer tilizers to Helianthus annuus .L is recommended to the far mers for attaining better growth, biochemical constituents and antioxidant enhancement.

Table 4: Effect of SLF of Sargassum wightii on antioxidant enzymes of the leaves of Helianthus annuus.L

45DAS

0.47±0.02

Control

30DAS

1 .52±0.005

1.90±0.07

1.02±0.01

2.88±0.06 (51.57) 2.90±0.05 (90.78) 1.71±0.06 (67.64)
- Values are expressed as mean ± SD DAS - days after sowing

Control

SLF (2.5%)

2.49±0.02 (31.05) 2.61±0.05 (70.71) 1.53±0.01 (50.0)

SLF (5.0%)

2.52±0.02 (32.63) 2.72±0.05 (78.94) 1.59±0.01 (55.88)

Chemical fertilizer

1.63±0.05

0.97±0.03

0.78±0.03 (65.95) 2.81±0.06 (72.39) 1.66±0.04 (71.13)

SLF (2.5%)

0.75±0.05 (59.57) 1.97±0.01 (20.85) 1.63±0.01 (68.04)

SLF (5.0%)

0.72±0.02 (53.19) 1.99±0.01 (22.08) 1.65±0.01 (70.10)

Chemical fertilizer

Atalase (Units/g protein) Peroxidase (Units/g protein) Glutathione Peroxidase

Parameters

SLF - Seaweed Liquid Fertilizer

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