www.thaiagj.org
Thai Journal of Agricultural Science 2014, 47(1): 45-50
Adsorption of Minerals on Epidermal Wall Layer of SLF
Treated Vigna mungo L.
G. Ganapathy selvam and K. Sivakumar*
Division of Algal Biotechnology, Department of Botany, Faculty of Science
Annamalai University, Annamalai Nagar-608602, Tamil Nadu, India
*Corresponding author, Email: [email protected]
Abstract
The experiment was conducted on Vigna mungo an important pulse crop of Tamil Nadu, South
India. It is to study the effect of the green seaweed Ulva reticulata as fertilizer on the growth of
black gram. The various concentrations of Seaweed liquid fertilizer from U. reticulata such as 1%,
2.5%, 5%, 7.5%, and 10% w/v was applied as foliar spray. Further epidermal peal from the leaf of
10% SLF treated Vigna mungo were subjected to scanning electron microscopy with energy
dispersive spectroscopy, it revealed that outer portion of epidermal wall contained ten elements in
the following order: Ca>P>Na>Mg>Zn>N>Fe>K>Si>S whereas the inner portion contained
eight elements in the following order: Mg>Mn>Na>N>Fe>Si>S>K. The present study revealed
that the seaweed Ulva reticulata foliar sprayed leaf epidermal portion occupied high amount of
mineral content. This practice could meet the manurial requirement for organic farming and serve
as a cost effective eco-friendly approach for sustainable agriculture and environment.
Keywords: Ulva reticulata, Seaweed Liquid fertilizer, Scanning electron microscope, Vigna mungo
Introduction
Seaweeds have rich source of minerals, especially
for human nutrition. The nutritional properties of
seaweeds are usually determined from their biochemical composition viz., protein, carbohytrates,
vitamins and amino acids (Darcy-Vrillon, 1993;
Mabeau and Fleurence, 1993). In recent years,
seaweed extracts are marketed as liquid fertilizer
since they contain many growth promoting
substances like auxins, gibberellins, trace elements,
vitamins, amino acids and micronutrients and can be
used as foliar spray to increase the amount of limiting
nutrients leading to higher biomass (Abetz, 1980).
Seaweeds may contain a high mineral content, as
their cell wall polysaccharides and proteins contain
anionic carboxyl, sulphate and phosphate groups that
are excellent binding sites for metal retention (Davis
et al., 2003). Mineral and chemical constituents of the
different species of marine algae are from Saurashtra
Coast (Karthikai Devi et al., 2009). Marine algae
have been consumed traditionally in Asia but only
occasionally in other parts of the world (Nisizawa et
al., 1993). From nutritional point of view, seaweeds
are characterized by high concentrations of fibre and
minerals (Burtin, 2003), low fat content and in some
cases relatively high protein levels (Galland-Irmouli
et al., 1999). Identification of organic sources for
seed and foliar treatment for yield increment will be
advantageous and the seaweeds become prominent
candidates since these marine resources in shallow
and deep water remain under-utilized for enhancing
crop production (Kaliaperumal et al., 2004). Much
work have been carried out on determining the effect
of SLF on black gram (Venkataraman Kumar and
Mohan, 1997); Dolichous (Anantharaj and
Venkatesalu, 2001). Considering the above important
facts, the present study was carried out to find out the
effect of SLF on the leaf area and distribution of
minerals using SEM-Energy dispersive spectroscopy
analysis. Turan and Kose (2004) on grapevine,
Mancuso et al. (2006) and Rathore et al. (2009) on
46
G. Ganapathy selvam and K. Sivakumar
soybean observed increases in yield as well as N, P
and K with application of seaweed extract. Gajewski
et al. (2008) on Chinese cabbage revealed that
application of Goteo (an organic mineral fertilizer
which contains algae extract Ascophyllum nodosum
with addition of phosphorus) increased yield,
marketable heads as well as vitamin C content
compared to the untreated cabbage whereas; slightly
higher nitrate content was noted. Zodape et al.
(2008), Arthur et al. (2003) on pepper and Zodape et
al. (2010) on mung bean indicated that application of
seaweed extract significantly increased seed yield and
pod weight as well as improved nutrional values of
seeds, i-e., protein and carbohydrates. Also,
Eyszkowska et al. (2008) reported that nitrate in
lettuce of examined cultivars insignificantly
increased after treatments with Goteo and
Aminoplant (an organic fertilizer which contains
aminoacids and short peptide chains). Abdel
Mawgoud et al., (2010) cleared that application of
seaweed extract at concentrations of 1, 2 and 3 g L-1
increased the response of all growth parameters and
yield of watermelon.
Energy dispersive spectroscopic analysis (EDS)
provides a unique approach for obtaining quantitative
compositional analysis of individual cell and intra
cellular compartments. Elemental quantification of
semi-thin sections with electron probe X-ray
microanalysis (EPMA) in generally based on the
linear relationship between elemental concentration
and the ratio of number of characteristic/continues
like X-ray photons (Shuman et al., 1976; Silverberg,
1976; Kitazawa et al., 1983). The macro and micro
elements are generally regarded as being cell wall
associated and has been detected by X-ray micro
analysis in a range of algal cells including blue green
algae (EI Bestway et al., 1996; Krivtso et al., 2000).
The amount of different chemical elements present in
the leaf of the Vigna mungo was determined using
Energy dispersive spectroscopy analysis in the
present study.
Materials and Methods
The seaweed used in the present study was Ulva
reticulata belonging to the class chlorophyceae. It
was collected from the coastal area of Tuticorin and
other places of South East Coast of Tamil Nadu (9º
Thai Journal of Agricultural Science
25’ N and 79º 15’ E) during April 2013. The algal
sample was handpicked and washed thoroughly
with seawater to remove all the impurities, sand
particles and epiphytes. It was kept in an ice box
containing slush ice, transported to the laboratory
and washed thoroughly using tap water to remove
the salt on the surface of the material. The water
was drained off and the algal material was spread
on blotting paper to remove excess water. Voucher
specimen of the U. reticulata was deposited in a
Herbarium in Department of Botany, Annamalai
University.
One kg of dry seaweed was cut into small pieces
and boiled with 1 liter of distilled water for an hour
and filtered. The filtrates were taken as 100%
concentration of the SLF and from this different
concentration (1%, 2.5%, 5%, 7.5% and 10%) were
prepared using distilled water (Bhosle et al., 1975).
As the SLF contained organic matter, the SLF was
refrigerated between 0ºC and 4ºC. The colour was
observed visually and the pH was measured using
pH meter. Calcium, Magnesium, Manganese,
Sodium, Potassium, Iron, Chloride, Sulphur, Zinc,
Copper and Nitrate content of the SLF were
analysed following method of American Public
Health Association (APHA, 1995).
The certified seeds of Vigna mungo were
procured from Regional Pulses Research Station,
Tamil Nadu Agricultural University, Coimbatore.
They were surface sterilized with 0.1 % mercuric
chloride and then sown in field. Experimental trail
was conducted at Botanical garden, Annamalai
University, Annamalainagar on Vigna mungo seed
was sown in field in 4m x 3m plot. One or two seed
were sown along a side of the ridges at 30 cm
spacing. For each experiment ten plants per row
was taken. Five treatments were given to the field
plants namely foliar spray of 1 %, 2.5%, 5%, 7.5%,
and 10% of aqueous seaweed extract. In each of the
foliar treatment, 100 mL aqueous extract was
applied. The first spray treatment was given to 15day-old seedlings. Thereafter, three sprays at
interval of 15 days each were given up to 60 days.
The control set was treated only with distilled water
as foliar spray. Growths parameters like shoot
length, root length and leaf area were recorded at
15th, 30th, 45th and 60th day.
Vol. 47, No. 1, 2014
Minerals adsorption of SLF treated vigna mungo L.
Epidermal Study
Epidermal peels were obtained uniformly from
the fourth leaf of the treated and control of pot
experiment plants by mechanical means and were
stained with aqueous safranin (1%). They were
mounted in 50% glycerin and sealed with DPX.
The variations in epidermal cells and stomata cells
among the treated and control was observed under
light microscope as well as Scanning Electron
Microscope (SEM).
Scanning Electron Microscope with EDS studies
Leaves of SLF treated V. mungo were fixed in
3% glutaraldehyde in 0.1 M phosphate buffer at
(pH 6.8). Specimens were dehydrated through a
graded series of alcohol for 12-15 min interval at
4C up to 70% of alcohol. Then dehydrated
specimens were treated with critical point drier
(CPD) and fixed on a stub and the specimens were
coated with silver and examined with Joel JSM56010 LV with INSA-EDS. Electron micrographs
were taken selectively from the computer screen.
Simultaneously, the selected electron micrographs
of leaf portion of adaxial layer from 2.5% SLF
treated and control plants were subjected to Energy
Dispersive Spectroscopic analysis. This was
conducted with an EDS 700 series interfaced with a
data general NOVA2 computer and a Texas
instrument silent 700 ASR. The EDS X-ray
spectrometer was interfaced with a scanning
electron microscope (20 KV) stage. The area of
different components such as epidermal portion and
cellular inclusion were analyzed. To find out the
fluxes, both the count per second (S-1 or CPS)
value and the apparent relative atomic percentage
of weight particular mineral in different
composition details were documented.
Results
Physico-chemical properties of SLF of U.
reticulata before preparation of different
concentrations were shown in Table 1. The growth
parameters such as shoot length, root length, and
leaf area were increased up to 2.5% SLF treatments
thereafter decreased up to 10% SLF treatments,
whereas control plants showed lowest value in all
growth parameters comparatively (Table 2).
Epidermal cells (Non-costal) of both surfaces were
47
Table 1 Physico-chemical analysis of Ulva reticulata
SLF.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
Parameters
Colour
pH
Calcium (mg L-1)
Magnesium (mg L-1)
Manganese (mg L-1)
Sodium (mg L-1)
Potassium (mg L-1)
Iron (mg L-1)
Phosphate (mg L-1)
Chloride (mg L-1)
Sulphate (mg L-1)
Zinc (mg L-1)
Copper (mg L-1)
Nitrate (mg L-1)
Values
Green
7.12
157.52
109.25
76.85
285.08
176.2
5.12
45.05
2230.36
56.42
1.95
1.75
126.85
large, sometimes small, irregularly distributed,
anisodiametric, sometime tending to became
isodiametric; walls thin and arched or slightly
sinuous, stomata are sub-circular to oval,
irregularly distributed, found on both surface
(epiamphistomatic) (Figures 1 and 2). Electron
micrograph of leaves shows upper and lower
epidermis, the epidermal cells are larger and
enclose stomata. Stomata are paracytic and present
in both the epidermal layers. The upper epidermis
shows less number of stomata were as lower
epidermis higher numbers. The present study
revealed that the seaweed Ulva reticulata 2.5%
foliar sprayed leaf epidermal portion occupied high
amount of mineral content. The results obtained
from the EDS analysis of V. mungo, different
chemical elements present in the cell wall of 2.5%
SLF treated leaf outer and inner portion of
epidermal wall were shown Figure 3 totally outer
portion of epidermal wall contained ten elements
namely Ca, P, Na, Mg, Zn, N, Fe, K, Si, and S were
observed. The inner portion contained eight
elements namely Mg, Mn, Na, N, Fe, Si, S and K
(Figure 4). The order of chemical elements from
epidermal wall outer and inner portion of the leaf of
SLF treated V. mungo as follows were
Ca>P>Na>Mg>Zn>N>Fe>K>Si>S and Mg>Mn>
Na>N>Fe>Si>S>K respectively.
48
G. Ganapathy selvam and K. Sivakumar
Thai Journal of Agricultural Science
Table 2 Effect of different concentration of Ulva reticulata seaweed liquid fertilizer on shoot, root length (cm) and leaf
area (mm2) in Vigna mungo L
SLF
Con.
(%)
Control
1
2.5
5
7.5
10
Shoot
length
13.72
0.08
12.80
0.09
16.25
0.08
16.05
0.12
15.16
0.09
14.05
0.09
15th day
Root
length
8.55
0.05
9.56
0.06
12.15
0.06
10.86
0.08
9.35
0.05
7.88
0.07
Leaf
area
18.34
0.09
25.95
0.16
36.85
0.19
34.94
0.21
22.76
0.11
21.15
0.14
Shoot
length
15.72
0.10
23.65
0.13
26.13
0.11
23.45
0.17
20.18
0.12
18.15
0.19
30th day
Root
length
13.63
0.07
15.66
0.95
15.80
0.08
16.18
0.08
14.05
0.07
14.18
0.09
Leaf
area
25.54
0.25
33.25
0.20
45.94
0.33
43.28
0.32
32.12
0.22
28.05
0.19
Shoot
length
25.32
 0.15
27.85
0.19
30.85
0.25
28.72
0.27
27.15
 0.14
26.25
0.15
45th day
Root
length
16.64
0.08
21.28
0.12
21.65
0.12
18.35
0.21
18.02
0.09
17.25
0.09
Leaf
area
38.15
0.22
43.75
0.26
53.38
0.36
50.34
0.34
44.17
0.26
39.05
0.26
Shoot
length
32.62
0.15
39.13
0.22
40.25
0.28
36.98
0.25
31.86
0.18
31.12
0.23
60th day
Root
Leaf
length area
20.14
39.24
 0.12  0.24
22.89
47.13
0.14
 0.27
25.65
56.68
0.17
 0.33
25.15
52.35
0.26
 0.26
22.35
46.55
0.13
 0.27
21.49
40.38
0.16
 0.35
Discussion
Figure 1 SEM image of leaf epidermal outer layer showing
stomata of V. mungo treated with 2.5% U. reticulata (SLF).
Figure 2 SEM image of leaf epidermal inner portion of V.
mungo treated with 2.5% U. reticulata (SLF).
The results were in agreement with the previous
studies on growth stimulation of SLF prepared
Gracilaria, Sargassum and Caulerpa on crops like
cumbu by (Murugalakshmi Kumari et al., 2003)
and cluster bean by (Thirumalthangam et al., 2003).
The seaweed liquid extract of Sargassum also
promoted the seedling growth of green gram and
black gram at the concentrations below 1.5%
(Venkataraman Kumar et al., 1993) macro and
micro elements (Cahallen and Hemingway 1965) S.
vulgare with 1.5% seaweed extract prepared from
H. clathratus (Ashok et al., 2004).
The energy dispersive X-ray microanalysis
provides a unique approach for obtaining
qualitative and quantitative compositional analysis
of individual cell and intra cellular compartment to
localize distribution of chemicals elements of leaf
differed not only by quality but also in quantity.
The high value of calcium obtained in the leaf is
understandable since its involvement in the
formation of cell wall layer. Magnesium (Mg)
present in the leaf because it forms nucleus for the
porphyrin ring and hence its presence in the
photosynthetic mechanism is understandable. The
cell structure and different cellular inclusions have
been reported by Sivakumar and Rangasamy (1999)
using x-ray microanalysis EDAX. Electron
microscopic studies and x-ray microanalysis on
Sargassum wightii were made by Sundari and
Selvaraj (2009). Ganapathy selvam and Sivakumar
Vol. 47, No. 1, 2014
Minerals adsorption of SLF treated vigna mungo L.
Figure 3 SEM image of leaf epidermal outer layer showing
stomata of V. mungo treated with 2.5% U. reticulata (SLF)
subjected EDS analysis.
49
reticulata. Thus one can prefer the seaweed extracts
of U. reticulata to increase crop productivity by
spraying 2.5% concentration. Promising increased
crop yield, nutrient uptake, resistance to frost and
stress, improved seed germination of reduced
incidence of fungal and inspect attack have been
resulted by application of SLF. These fertilizers
harness atmospheric nitrogen and make it directly
available to the plants. They increase the
phosphorous content of the soil by solubilizing and
releasing unavailable phosphorous. Bio-fertilizers
improve root proliferation due to the release of
growth promoting hormones. Hence use of modern
agriculture in conjunction with traditional farming
practices is the sustainable solution for the future.
Acknowledgments
The authors are thankful to Head of the
Department of Botany, Annamalai University for
having provided laboratory facilities.
References
Figure 4 SEM image of leaf epidermal inner portion of V.
mungo treated with 2.5% U. reticulata (SLF) subjected EDS
analysis.
(2012) reported the effect of SLF of Ulva reticulata,
as biochemical characteristics of Vigna mungo as
well as leaf morphometric analysis such as epidermal
and stomata cell variation and distribution of
minerals in the leaf.
The data generated from study reveal that SLF
of H. musciformis could be used as foliar spray at
low concentration of 2% to maximize the growth
and yield of A. hypogea and also increase the
number of stomata in the leaf (Ganapathy selvam
and Sivakumar 2014). This practice could meet the
manurial requirement for organic farming and serve
as a cost effective eco-friendly approach for
sustainable agriculture and environment.
Conclusions
In conclusion, the observations on SLF treated
V. mungo plants suggested that growth parameters
characteristics of pulse crop plants might be
promoted by micro and macro elements and growth
promoting hormones present in the extract of Ulva
Abdel-Mawgoud, A.M.R., A.S. Tantawy, M. Magda Hafez
and A.M. Habib. 2010. Seaweed extract improves
growth, yield and quality of different watermelon
hybrids. Res. J. Agric. Biol. Sci. 6: 161-186.
Abetz, P. 1980. Seaweed extracts: Have they a placed in
Australian Agriculture or Horticulture?. J. Aus. Inst.
Agric. Sci. 46: 23-29.
Anantharaj, M. and V. Venkatesalu. 2001. Effect of
seaweed liquid fertilizer on Vigna catajung. Seaweed
Res. Utiln. 23: 33-39.
APHA. 1995. Standard Methods for Examination of Water
and Waste Water Analysis, 19th ed., Washington.
Arthur, G.D, W.A. Stirk and Van J. Staden. 2003. Effect
of seaweed concentrates on the growth and yield of
three varieties of Capsium annuum, South Afr. J.
Bot. 69: 207-211.
Ashok, V., N. Vijayanand and S. Rathinave. 2004. Biofertilizing efficiency of seaweed liquid extract of
Hydroclathrus clathratus on Sorghum vulgare,
Seaweed Res. Utiln. 26: 181-186.
Bhosle, N.B, V.K. Dhargalkar and A.G. Untawale. 1975.
Effect of seaweed extract on the growth of Phaseolus
vulgaris L. Indian J. Mar. Sci. 4: 207-210.
Burtin, P. 2003. Nutritional value of seaweeds. EJEAF
Che. 2: 498-503.
Cahallen, S.B. and J.G. Hemingway. 1965. Growth of
higher plants in response to feeding with seaweed
extract. Proc. Fifth Int. Seaweed Symp. 5: 359-367.
Darcy-Vrillon, B. 1993. Nutritional aspects of the
developing use of marine macroalgae for the human
food industry. Int. J. Food Sci. Nutr. 44: 23-35.
50
G. Ganapathy selvam and K. Sivakumar
Davis, T.A., B. Volesky and A. Mucci. 2003. A review of
the biochemistry of heavy metal biosorption by brown
algae. Water Research 37: 4311-4330.
EI-Bestway, E., E. Bellinger and D.C. Sigee. 1996.
Elemental composition of phytoplankton in a
subtropical lake: X-Ray microanalytical studies on the
dominant algae of Spirulina and Cyclotella. Eur. J.
Phyco. 31: 157-166.
Eyszkowska, M., J. Gajc-Wolska and K. Kubi. 2008. The
influence of biostimulators on yield and quality of leaf
and iceberg lettuce grown under field conditions, 2834. Conferences of Biostimulators in Modern
Agriculture “Vegetable Crops”, Warsaw.
Gajewski, M., G. Katarzyna and J. Bobruk. 2008. The
influence of Goemar Goteo biostimulator on yield and
quality of two Chinese cabbage cultivars, 23-27.
Conferences of Biostimulators in Modern Agriculture
“Vegetable Crops”, Warsaw.
Galland-Irmouli, A.V., J. Fleurence, R. Lamghari, M.
Rouxel, C. Barbaroux, O. Bronowicki and J.P.
Villaume. 1999. Nutritional value of proteins from
edible seaweed Palmaria palmata (Dulse). J. Nutri.
Biochem. 10: 353-359.
Ganapathy Selvam, G. and K. Sivakumar. 2013. Effect of
Foliar spray from Seaweed Liquid Fertilizer of Ulva
reticulata (Forsk.) on Vigna mungo L. and their
elemental composition using SEM–Energy Dispersive
Spectroscopic analysis. A. Paci. J. Reprod. 2: 119-125.
Ganapathy Selvam, G. and K. Sivakumar. 2014. Influence
of seaweed extract as an organic fertilizer on the
growth and yield of Arachis hypogea L. and their
elemental composition using SEM–Energy Dispersive
Spectroscopic analysis. A. Paci. J. Reprod. 3: 18-22.
Hayat, M.A. 1970. Principles and Techniques of Electron
Microscopy: Biol Appli 1, Reinhold Co., New York
and London.
Kaliaperumal, N., S. Kalimuthu and J.R. Ramalingam.
2004. Present scenario of seaweed exploitation and
industry in India. Seaweed Res. Utiln. 26: 47-53.
Karthikai Devi G., G. Thirumaran, K. Manivannan and P.
Anantharaman. 2009. Element Composition of Certain
Seaweeds from Gulf of Mannar Marine Biosphere
Reserve; Southeast Coast of India. World J. Dairy and
Food Sci. 4: 46-55.
Kitazawa, T.H., A.V. Somlyo and A.P. Somlyo. 1983.
Quantitative electron probe analysis; problems and
solutions. Ultramicroscopy 11: 251-262.
Krivtso, V., E. Bellinger and D. Sigee. 2000. Incorporation
of the intercellular elemental correlation pattern in to
simulation model of phytoplankton uptake and
population dynamic. J. Appl. Phycol. 1: 53-59.
Mabeau, S. and J. Fleuraence. 1993. Seaweeds in food
products, biochemical and nutritional aspects. Trends
Food Sci. Technol. 4: 103-107.
Mancuso, S., E. Azzarello, S. Mugnai and X. Briand. 2006.
Marine bioactive substances (IPA extract) improve
Thai Journal of Agricultural Science
foliar ion uptake and water tolerance in potted Vitis
vinifera plants. Adv. Hort. Sci. 20: 156-161.
Murugalakshmi Kumari, R., V. Ramasubramanian and K.
Muthuchezhian. 2003. Studies on utilization of
seaweed as an organic fertilizer on the growth and
some biochemical characteristics of blackgram and
cumbu. Seaweed Res. Utiln. 24: 125-128.
Nisizawa, K. 1993. Monthly determination of alginate,
M/G ratio, mannitol and minerals in cultivated
Laminaria japonica. Nippon Suisan Gakkaishi 59: 295299.
Rathore, S.S., R. Chaudhary, G.N. Boricha, A. Ghosh, B.
P. Bhatt, S.T. Zodape and J.S. Patolia. 2009. Effect of
seaweed extract on the growth, yield and nutrient
uptake of soybean (Glycine max) under rainfed
conditions. S.A.J. Bot. 75: 351-355.
Shuman, H. A.V. Somlyo and A.P. Somlyo. 1976.
Quantitative electron probe microanalysis of biological
thin sections methods and validity. Ultra Microscopy 1:
317-339.
Silverberg, B.A. 1976. Cadmium-induced ultra-structural
changes in mitochondria of fresh water algae.
Phycologia 15: 155-159.
Sivakumar, K. and R. Rangasamy. 1999. X-ray
microanalysis of cell wall, plastids and Floridian starch
of Hypnea musciformis (Wulf) Lamour. (Gigartinales,
Rhodophyta). Seaweed Res. Utiln. 21: 49-53.
Sundari, K. and R. Selvaraj. 2009. Electron microscopic
study and X-Ray microanalysis of Sargassum species.
Seaweed Res. Utiln. 3: 85-94.
Thirumalthangam, R., M.V. Rani and S.P. Marian. 2003.
The Effect of seaweed liquid fertilizers on the growth
and biochemical constituents of Cyamopsis
tetragonoloba (L.) Taub. Seaweed Res. Utiln. 25: 99103.
Turan, K. and M. Kose. 2004. Seaweed extract improve
copper uptake of Grapevine (Vitis vinifera). Soil Plant
Sci. 54: 213-220.
Venkataraman, K.V., V.R. Mohan, R. Murugeswari and
M. Muthusamy. 1993. Effect of crude and commercial
seaweed extracts on seed germination and seedling
growth in green gram and black gram. Seaweed Res.
Utiln. 16: 23-27.
Venkataraman, K.V. and V.R. Mohan. 1997. Effect of
seaweed extracts SM3 on the cyanobacterium,
scytonema species. Seaweed Res. Utiln. 19: 43-47.
Zodape, S.T., S. Mukhopadhyay, K. Eswaran, M.P. Reddy
and J. Chikara. 2010. Enhanced yield and nutritional in
green gram (Phaseolus radiata L.) treated with
seaweed (Kappaphycus alvarezii) extract. J. Sci. Indus.
Res. 69: 468-471.
Zodape, S.T., V.J. Kawarkhe, J.S. Patolia and A.D.
Warade. 2008. Effect of liquid seaweed fertilizer on
yield and quality of okra (Abelmoschus esulentus L.) J.
Sci. Indus. Res. 67: 1115- 1117.
Manuscript received 12 April 2014, accepted 29 November 2014
Now online at http://www.thaiagj.org