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INFLUENCE OF CHLORELLA VULGARIS ON THE GROWTH, BIOMASS AND

YIELD PARAMETERS OF MAIZE

Article · September 2017

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 INFLUENCE OF CHLORELLA VULGARIS ON THE GROWTH, BIOMASS AND YIELD
PARAMETERS OF MAIZE

SAMBO, G.Y.*, CHIA, M.A. AND IORTSUUN, D.N.

Department of Botany, Ahmadu Bello University, Nigeria

ABSTRACT
The potential of using the freshwater microalga Chlorella vulgaris as a biofertilizer for maize (Zea mays L. Var
Sampea 8) cultivation was investigated. Different concentrations of C. vulgaris biomass concentrations, 0.5, 1.0
and 1.5 g FW representing 0.5, 1.0 and 1.5% w/v water, respectively, were added to 3 Kg soil per pot before
sowing maize seeds (pre-application), and after germination (post-application). Under each post-application
experiment, the different biomass concentrations were combined with and without N.P.K. chemical fertilizer at
50 and 100% the recommended dose. In general, the pre-application of C. vulgaris resulted in higher growth,
chlorophyll content, biomass production and yield parameters, than the post-application. The combination of C.
vulgaris with half the recommended dose of chemical fertilizer increased growth, crop yield and chlorophyll
contents of maize, than only pre- or post-application of the microalga with 100% or without chemical fertilizer.
Protein, carbohydrate and fiber content of maize seeds grown with 1.5% C. vulgaris was not significantly
different from those grown with full dose chemical fertilizer. The results of the present study show that the
combination of C. vulgaris treatment with 50% chemical fertilizer may provide a possible means of reducing
chemical fertilizer usage without compromising growth and yield.

Keyword: Fertilizer, growth, microalgae, nutrients, yield

*Corresspondence: [email protected]

INTRODUCTION
Soils are simultaneously exposed to various parameters of crop plants [7, 8]. Among microalgae
pressures in the form of erosions, loss of nutrient, used as biofertilizers, cyanobacteria has drawn most
accumulation of salts and toxic elements, water of the attention due to their ability to fix
logging and un-balanced nutrient compensation, atmospheric nitrogen and stabilize soils, thereby
which determines their suitability for farming [1]. encouraging high vegetative and reproductive
The excessive use of inorganic/chemical fertilizers growth in crop plants [3]. Unfortunately, very little
have led to adverse environmental, agricultural and is known about the role of green algae as
health consequences such as soil erosions, loss of biofertilizer on plants. However, few studies
nutrient, accumulation of salts and other toxic support the use of green algae as biofertilizer
elements, water logging e.t.c. As alternatives, because of their high biochemical content. For
organic wastes and bio-fertilizers sources can example, C. vulgaris contains high amounts of
provide the nutrient requirement of crops in a more macro and micronutrients in the form of its
eco-friendly manner [2]. constituents or metabolites such as carbohydrates,
For sustainable agriculture, bio-organic lipids, vitamins, amino acids, polypeptides,
farming is encouraged to limit the deleterious antibacterial and antifungal substances, and growth
effects of using inorganic fertilizers. This leads to promoting hormones like cytokinins [9-14].
the production of safer agricultural products for The use of microalgal fertilizer improves
human and animal consumption, as well as soil microbial activities and health compared to
enhancing the biological control of pests and soil soils treated with inorganic fertilizers [15]. A novel
borne pathogens [3]. A biofertilizer is an organic approach has been the application of bio-filmed
substance which contains living microorganisms biofertilizers with low levels of chemical fertilizers
that colonize the rhizosphere when applied to the to reduce the amount of chemical fertilizers used in
soil, and promotes plant growth by increasing the farming [16]. These applications have given yields
supply or availability of essential nutrients to the comparable to those of 100% fertilizers in tea, rice
host plant [4]. Biofertilizers add nutrients through and maize [17, 18, 19]. However, to the best of our
the natural processes of nitrification, solubilizing knowledge, nothing is known about the potential of
phosphorous and stimulating plant growth through using C. vulgaris alone or in combination with
the synthesis of phytohormones [5]. A number of chemical fertilizers for different crop plants.
microflora have been identified for their potential Furthermore, the applicability of biofertilizers in
as fertilizers due to their role in primary production, different seasons is generally not known. Tropical
nutrient balancing, pathogen suppression and regions of the world have characteristic rainy and
maintenance of soil structure and water dry seasons, and irrigation farming is usually
relationships [6]. The use of algal extracts as carried out in the dry months [20]. This increases
biofertilizers is encouraged for increasing growth the amount of chemical fertilizers applied to the

Nigerian Journal of Scientific Research, 16 (5): 2017; September- October; njsr.abu.edu.ng 624
Sambo et al. (2017); Chlorella vulgaris as a biofertilizer for maize

environment, and the risk of environmental vulgaris biomass concentrations were combined
contamination and degradation. In addition, with 50 and 100% of the recommended dose (120
environmental conditions during each season vary, Kg per hectare, corresponding to 3 g per
and results obtained in one season or in the experimental bag) of chemical fertilizer [120 kg N,
laboratory may not be comprehensive enough to 60 kg P2O5 and 60 kg K2O]. No inorganic fertilizer
accommodate year round changes in field or C. vulgaris was added to the control treatments.
conditions. Therefore, the objective of this study Three maize seeds were planted per replicate of
was to investigate the potential of C. vulgaris as a each treatment at a depth of 2-3 cm per bag.
biofertilizer. Planting took place under natural environmental
conditions of light and temperature. All
MATERIALS AND METHODS experimental treatments were carried out in
triplicates.
Algal culture conditions
Chlorella vulgaris UTEX2714 was obtained from Growth measurement
the UTEX Culture Collection of Algae, The Shoot length of maize was measured from 10 days
University of Texas at Austin, and used for the after germination, and subsequently, on a weekly
biofertilization experiments. Stock cultures were basis for six weeks. Fresh and dry weights were
maintained in Bold Basal Medium (BBM) using a determined gravimetrically. For dry weight
semi-continuous batch system as described in [12, measurements, plant materials were oven at 40°C
13]. Culture medium sterilization was performed by for 3 days, and weighed to the nearest mg. Number
autoclaving at 121oC for 30 min. Cultures were of leaves per plant and leaf area index
kept under continuous controlled conditions of light measurements were performed following the
intensity (150 μmol m-2 s-1) and temperature (23+ 2 procedures described in [21]. At the end of the
°C). The microalga was cultured in a 20 L airlift cultivation period (87 days), all fruited maize plants
photobioreactor containing 15 L of BBM medium. were harvested. Shoots and fruits were separated
and stored in paper bags, and yield parameters such
Source of maize seeds and soil preparation as weight of maize cob, number of seed rows per
Maize seeds (variety SAMMAZ 12) were obtained plant, and cob length were determined.
from the SEED UNIT of the Institute for
Agricultural Research, Ahmadu Bello University Chemical analysis
(ABU), Zaria. Maize seed planting was done in the Chlorophylls were extracted by grinding 0.5 g of
Biological Science Garden, ABU, in loamy soil maize leaf in 10 mL 80% acetone. The amount of
containing equal amount of clay and silt. The soil chlorophyll a, chlorophyll b, total chlorophyll and
mixture was packed into polythene bags of 25 by chlorophyll stability index present in the extract
25 cm containing 3kg of soil. was calculated [22]. Analysis of total carbohydrates
of the harvested seeds was performed according to
Experimental design the modified phenol-sulfuric acid technique [23]
Microalgal biomass was obtained by low speed using glucose as standard. Total fiber and total
centrifugation (1500 g) of exponential phase protein concentrations were determined from 2g of
growing C. vulgaris, which was then rinsed three harvested seeds per treatment [24]. Protein content
(3) times and re-suspended in sterilized distilled was determined by using nitrogen to protein
water to remove traces of growth medium [17]. All conversion factor of 5.75. Lipid content was
pre-planting application (pre-application) and post- extracted and measured from 2 g of seed samples
planting application (post-application) experiments [25]. Total nitrogen (N) and phosphorus (P)
were carried out in three months of dry (November concentrations in soil, microalga and harvested
2013 – January 2014) and three months of rainy maize seeds were determined using the Kjeldahl’s
(August – October 2013) seasons. In the dry method [26, 27]. The concentrations of available
season, watering was done every other day using potassium (K), copper (Cu), manganese (Mn), zinc
500 mL of distilled water per experimental bag to (Zn) and iron (Fe) in the soils with or without
ensure the plants do not suffer desiccation. In the chemical fertilizer and C. vulgaris were determined
pre- application experiment, 0.5, 1, and 1.5 g FW of using Atomic Absorption Spectroscopy (AAS)
the microalga were re-suspended in 100 mL water model number LAAS-210 [27] after harvest.
to give 0.5, 1, and 1.5 % w/v, respectively, and
added to the soil 15 days prior to planting. On the Data analysis
other hand, in the post-planting application The data obtained were subjected to Levene’s
experiments, inorganic fertilizer and C. vulgaris homogeneity of variance test. Subsequently, the
were applied separately and in combination, 10 data were subjected to factorial analysis of variance
days after planting and 4 weeks after first chemical (ANOVA). Significantly different means were
fertilizer and C. vulgaris application. Under separated using Duncan Multiple Range Test
combined treatment conditions, the different C. (DMRT).

Nigerian Journal of Scientific Research, 16 (5): 2017; September- October; njsr.abu.edu.ng 625
Sambo et al. (2017); Chlorella vulgaris as a biofertilizer for maize

RESULTS treatments were statistically significant (p<0.05) in

both wet and dry months. The control had the
In relation to the control, there was a significant lowest cob length, cob width and number of seed
increase (P<0.05) in N, P, K, Mn, Fe, Zn and Cu rows in maize. Although the pre and post-
concentrations in the soil with the application of C. application of 0.5% C. vulgaris did not cause
vulgaris and NPK fertilizer (Table 1-2). However, significantly different growth parameters from the
there was no significant difference (P<0.05) in the control in the dry season, changes in number of
concentration of Mn and Fe in C. vulgaris and NPK seed rows and cob length were significant (p<0.05)
fertilizer treated soils. Soils treated with NPK in the presence of C. vulgaris in both seasons in
fertilizer had the highest N, P, K, Mn, Zn, Fe and maize (Table 5).
Cu concentrations, followed by those inoculated Plant height, leaf area and leaf number
with C. vulgaris. increased in both seasons when the maize plants
Growth and crop yield parameters results were exposed to combined C. vulgaris and NPK
of maize exposed to different concentrations of fertilizer treatments compaed to the control (Table
NPK fertilizer and C. vulgaris treatments are shown 3). The combination of C. vulgaris (1.5%) and
in Tables 3-5. With increasing C. vulgaris biomass, 100% NPK fertilizer gave higher plant height, fresh
plant height, leaf area, number of leafs, fresh and dry weight, cob length, cob width and number
weight, dry weight, cob length, cob width, number of seed rows per plant, than their individual effects
of seed rows were increased in both dry and wet in both wet and dry months. The 100% NPK
months. Pre-application of C. vulgaris resulted in fertilizer treatment gave the highest leaf area and
higher plant height, leaf area, fresh weight, dry leaf number in both wet and dry months, while the
weight, cob length, cob width, and number of seed control had the lowest fresh and dry weight in both
rows per maize plant than the post-application of C. seasons. In addition, the 100% NPK fertilizer
vulgaris without chemical fertilizer in both rainy treatment had the highest cob length, cob width and
and dry seasons. The variations in growth and yield number of seed rows in maize in dry season.
parameters recorded between the C. vulgaris

Table 1: Nitrogen, Phosphorus and Potassium concentrations in Chlorella vulgaris and soil treated with
Chlorella vulgaris (1.5%) and chemical fertilizer at 100%.
Treatment N (%) P (µg/g) K (cmol/kg)

Soil 0.108±0.004d 14.65±2.81d 0.47±0.002c

Soil + 1.5% Chlorella 0.424±0.088c 52.76±0.95c 0.78±0.003bc

Soil + 100% NPK fertilizer 1.721±0.091b 230.00±2.33b 4.63±0.01b

C. vulgaris 6.226±0.026a 1570.00±0.01a 25.37±1.80a

Means having the same letter(s) per column are not significantly different (p>0.05).

Table 2: Micronutrients in the soil with application of Chlorella vulgaris and NPK fertilizer.
Treatment Mn (µg/g) Zn (µg/g) Fe (µg/g) Cu (µg/g)
Control 5.220±0.940b 280±0.190c 22.585±0.134b 0.215±0.005c

C. vulgaris 10.495±0.275a 3.470±0.230b 28.430±0.100a 0.260±0.010b

NPK fertilizer 11.030±0.300a 4.365±0.005a 28.295±0.075a 0.305±0.005a

Means having the same letter(s) per column are not significantly different (p>0.05).

The pre-application of 1.5% of Chlorella vulgaris total chlorophyll and chlorophyll stability index
and the combination of C. vulgaris and NPK were highest in pre-application treated maize plants
fertilizer resulted in higher levels of chlorophyll a, in both rainy and dry months, compared to the post-
b, total chlorophyll and chlorophyll stability index application treatments. The combination of C.
of maize in both dry and wet months, but showed vulgaris and chemical fertilizer resulted in an
no significant difference with pre-application of increase in chlorophyll b, total chlorophyll and
1.5% C. vulgaris in chlorophyll a in the dry months chlorophyll stability index production in both rainy
compared to the control (Table 6). The pre- and dry months than their individual effects.
application of C. vulgaris resulted in higher total Combination of C. vulgaris and chemical fertilizer
chlorophyll and chlorophyll stability index than its showed significance in chlorophyll a, b, total
post-application in rainy months. Chlorophyll b,

Nigerian Journal of Scientific Research, 16 (5): 2017; September- October; njsr.abu.edu.ng 626
Sambo et al. (2017); Chlorella vulgaris as a biofertilizer for maize

chlorophyll and chlorophyll stability index from the 100% chemical fertilizer showed that there was no
control. Proximate analyses of maize seeds of significant difference in total carbohydrate, fibre,
plants that were separately exposed to 1.5% C. lipid and protein production (Table 7).
vulgaris (pre-application and post-application) and

Table 3: Plant height, leaf number and leaf area of Zea mays treated with different pre and post-sowing
application of Chlorella vulgaris and NPK fertilizer concentrations during the rainy and dry seasons.
Treatment Plant height Leaf Leaf area (cm2)
(cm) number
Rainy Dry Rainy Dry Rainy Dry
Control 35.1±5.8dc 36.9±6.2c 6.9±0.9dc 7.5±0.9bc 57.9±11.7g 58.3±17.3g
Pre-application
0.5% CV 35.1±7.1dc 38.8±6.6dc 7.9±1.2c 7.8±0.7bc 81.2±33.0e 51.4±13.4h
cd d bc ab e
1.0% CV 44.3±7.6 44.6±6.9 8.3±1.4 9.0±1.0 97.2±29.2 75.0±19.4f
c cd b a d
1.5% CV 51.8±8.8 52.7±9.5 8.9±1.3 9.8±1.2 140.6±40.7 92.7±22.8e
Post-application
0.5% CV 36.0±5.9dc 36.5±5.9c 7.8±1.2c 8.3±0.5b 72.8±15.3f 54.1±12.1g
d d c b e
1.0% CV 40.7±7.7 43.5±7.2 8.1±1.2 8.5±0.8 90.1±22.5 70.0±15.6f
cd cd bc b d
1.5% CV 46.6±9.6 49.4±9.9 8.5±1.2 8.7±0.8 137.7±38.9 110.3±23.4e
bc c b b c
50% CF + 63.3±15.6 66.6±13.6 9.0±1.3 8.3±0.7 233.9±57.7 194.6±50.3d
0.5% CV
50% CF + 67.8±17.2bc 76.1±16.0bc 8.9±1.3b 10.1±0.9a 246.7±58.3bc 236.1±50.1c
1.0% CV
50% CF + 75.7±19.7b 82.5±18.4b 9.4±1.3ab 9.6±1.1ab 277.3±61.5b 271.2±54.8b
1.5% CV
100% CF + 80.8±18.8ab 92.5±20.1ab 9.0±1.5.0b 9.6±1.2ab 241.2±50.7bc 250.3±56.4bc
0.5% CV
100% CF + 82.8±19.5ab 93.8±20.4ab 9.3±1.4ab 10.1±1.2a 252.7±48.9bc 280.9±62.8ab
1.0% CV
100% CF + 86.7±20.3a 95.4±22.0a 9.6±1.4a 10.1±1.1a 316.5±63.5ab 297.5±65.6a
1.5% CV
50% CF 77.3±19.9b 85.9±18.5b 9.2±1.3ab 9.6±1.2ab 258.9±50.2b 256.7±55.9bc
ab ab a a a
100% CF 82.5±22.3 93.7±20.9 9.8±1.4 10.2±1.0 329.9±68.8 298.7±63.9a
Values are means of 3 replicates+ S.E.CV=Chlorella vulgaris, CF=chemical fertilizer. Means having the same letter(s) per column are not
significantly different (p>0.05).

Table 4: Mean dry and fresh weight of Zea mays treated with different Concentrations of Chlorella vulgaris and
NPK fertilizer during the rainy and dry seasons.
Treatment Fresh Weight (g) Dry weight (g)
Rainy Dry Rainy Dry
Control 138.1±2.4fg 121.27±4.47e 43.4±3.1ef 40.60±4.69de
Pre-application
0.5% CV 149.7±3.6f 137.02±1.01e 48.6±2.3e 46.83±3.54de
e d d
1.0% CV 211.8±2.0 193.77±3.27 64.6±3.7 64.22±8.45cd
de cd c
1.5% CV 244.6±2.7 215.90±9.07 87.1±1.5 77.50±2.56cd
Post-application
0.5% CV 128.8±2.9g 132.17±1.75e 46.6±2.8e 46.13±1.58de
ef de de
1.0% CV 176.3±2.0 152.83±1.73 55.8±3.3 52.85±1.65d
e cd cd
1.5% CV 223.8±0.7 210.400±2.96 81.1±2.0 73.80±1.47cd
d c bc
50% CF + 0.5% CV 319.4±2.5 292.11±5.37 108.7±2.4 94.16±4.86c
cd bc ab
50% CF + 1.0% CV 341.4±3.5 322.70±1.34 135.8±5.8 113.57±1.93bc
bc b ab
50% CF + 1.5% CV 368.0±4.3 363.80±3.87 140.4±2.5 121.80±2.76b
b a b
100% CF + 0.5% CV 411.4±2.8 405.70±2.63 130.9±4.0 130.40±9.61ab
ab a b
100% CF + 1.0% CV 427.9±5.5 411.20±6.97 133.6±2.2 136.10±9.81ab
a a a
100% CF + 1.5% CV 439.2±3.5 417.30±7.84 151.3±3.7 141.20±6.72a
c ab ab
50% CF 351.7±2.8 381.40±3.15 140.1±4.0 125.90±2.49ab
ab a a
100% CF 426.2±2.6 414.90±5.59 150.1±2.4 137.00±1.36ab
Values are means of 3 replicates+ S.E.CV=Chlorella vulgaris, CF=chemical fertilizer. Means having the same letter(s) per column are not
significantly different (p>0.05).

Nigerian Journal of Scientific Research, 16 (5): 2017; September- October; njsr.abu.edu.ng 627
 Sambo et al. (2017); Chlorella vulgaris as a biofertilizer for maize

Table 5: Mean crop yield of Zea mays treated with different Chlorella vulgaris concentration and NPK fertilizer
during the rainy and dry seasons.
Treatment Cob length (cm) Cob width (cm) Number of seeds row
Rainy Dry Rainy Dry Rainy Dry
Control 2.2±2.3d 0.0±0.0cd 0.8±0.8bc 0.0±0.0ab 0.0±0.0bc 0.0±0.0c
Pre-application
0.5% CV 3.0±3.0cd 0.0±0.0cd 1.4±1.4b 0.0±0.0ab 2.7±2.7b 0.0±0.0c
b bc a a a
1.0% CV 10.6±0.6 10.2±0.3 4.1±0.2 3.9±0.1 13.0±0.6 6.7±0.3b
b bc a a a
1.5% CV 11.0±0.2 10.3±0.2 4.2±0.2 4.2±0.1 15.0±1.0 7.7±0.7b
Post-application
0.5% CV 2.7±2.7d 0.0±0.0cd 0.8±0.8bc 0.0±0.0ab 0.0±0.0bc 0.0±0.0c
cd c b ab b
1.0% CV 4.8±2.6 1.8±1.8 1.4±1.4 0.7±0.7 3.7±3.7 0.0±0.0c
c c ab ab b
1.5% CV 6.5±3.3 3.2±3.2 2.8±1.4 1.2±1.2 4.7±2.6 1.3±1.3bc
b b a a ab
50% CF + 0.5% 14.0±0.4 14.0±0.4 4.4±0.0 4.4±0.1 13.7±0.8 13.7±0.9ab
CV
50% CF + 1.0% 16.1±0.9ab 16.7±0.3ab 4.5±0.3a 4.6±0.3a 14.0±0.0a 14.3±0.3ab
CV
50% CF + 1.5% 16.3±0.9ab 16.2±0.3ab 4.6±0.0a 4.4±0.4a 14.3±0.3a 14.0±2.0ab
CV
100% CF + 17.1±0.6a 16.3±0.4ab 4.1±0.1a 4.8±0.4a 13.3±0.7a 15.0±0.0a
0.5% CV
100% CF + 17.1±1.0a 16.9±0.2ab 4.1±0.2a 5.0±0.5a 13.7±0.9a 15.0±0.6a
1.0% CV
100% CF + 18.2±0.4a 17.1±0.5ab 4.5±0.2a 5.3±0.6a 14.7±0.3a 15.3±0.7a
1.5% CV
50% CF 15.5±0.4ab 15.4±0.5ab 3.7±0.2a 4.1±0.3a 11.0±1.0a 12.3±1.2b
a a a a a
100% CF 18.1±0.3 17.5±0.5 4.6±0.1 4.8±0.3 14.0±0.0 15.3±0.7a
Values are means of 3 replicates+ S.E.CV=Chlorella vulgaris, CF=chemical fertilizer. Means having the same letter(s) per column are not
significantly different (p>0.05).

Table 6: Mean chlorophyll content of Zea mays treated with different Concentrations of Chlorella vulgaris and
NPK fertilizer during the rainy season.

Treatment Chlorophyll a (mg/g) Chlorophyll b Total chlorophyll CSI

(mg/g) (mg/g)
Rainy Dry Rainy Dry Rainy Dry Rainy Dry
Control 0.8±0.0ab 0.8±0.0ab 0.8±0.2b 0.8±0.1c 1.6±0.2b 1.6±0.1b 100.0±1.0c 100.0±7.6bc
Pre-application
0.5% CV 0.8±0.1ab 0.8±0.0ab 0.8±0.1b 0.9±0.1bc 1.6±0.2b 1.7±0.2b 92.4±11.3cd 99.8±9.1bc
1.0% CV 0.8±0.0ab 0.9±0.0a 0.9±0.1b 0.7±0.1d 1.7±0.1b 1.9±0.3ab 103.5±7.1bc 91.0±2.0c
1.5% CV 0.9±0.0a 0.9±0.0a 1.1±0.1ab 1.0±0.1b 2.0±0.1ab 1.9±0.1ab 120.6±3.5a 111.7±4.1ab
Post-application
0.5% CV 0.8±0.0ab 0.8±0.0ab 0.9±0.2b 0.7±0.1cd 1.6±0.2b 1.5±0.1bc 95.6±13.5cd 96.3±7.6c
1.0% CV 0.8±0.0ab 0.8±0.1ab 0.8±0.1b 0.6±0.1de 1.6±0.1b 1.4±0.1bc 94.5±6.0cd 79.2±6.8cd
1.5% CV 0.8±0.0ab 0.9±0.0a 0.9±0.1b 0.9±0.1bc 1.8±0.1b 1.6±0.1b 104.0±8.4bc 106.9±8.7bc
50% CF + 0.8±0.4ab 0.8±0.0ab 1.0±0.1ab 1.0±0.1b 1.6±0.2b 1.9±0.0ab 110.5±7.9b 112.7±4.8ab
0.5% CV
50% CF + 0.9±0.0a 0.9±0.0a 1.0±0.1ab 1.1±0.1ab 1.9±0.1ab 1.9±0.1ab 110.3±2.5b 110.2±5.9ab
1.0% CV
50% CF + 0.9±0.0a 0.9±0.0a 1.2±0.0a 1.2±0.0a 2.1±0.0a 2.0±0.0ab 117.0±4.4ab 118.6±2.1a
1.5% CV
100% CF + 0.9±0.0a 0.8±0.0ab 1.2±0.0a 1.2±0.0a 2.0±0.0ab 2.1±0.0a 109.3±1.9b 114.6±1.2ab
0.5% CV
100% CF + 0.8±0.0ab 0.9±0.0a 1.2±0.1a 1.2±0.0a 2.0±0.1ab 2.1±0.0a 112.6±2.1b 113.3±2.3ab
1.0% CV
100% CF + 0.9±0.0a 0.9±0.0a 1.2±0.0a 1.2±0.0a 2.1±0.0a 2.1±0.0a 116.0±3.6ab 118.3±2.5a
1.5% CV
50% CF 0.8±0.0ab 0.9±0.0a 1.2±0.0a 1.2±0.1a 2.0±0.0ab 2.1±0.1a 116.4±1.7ab 117.5±3.4ab
100% CF 0.9±0.0a 0.8±0.0ab 1.2±0.0a 1.2±0.0a 2.1±0.1a 2.1±0.0a 119.5±3.4a 119.7±1.5a
Values are means of 3 replicates+ S.E. CF represents chemical fertilizer. Means having the same letter(s) per column are not significantly
different(p>0.05). *CSI= chlorophyll stability index

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Sambo et al. (2017); Chlorella vulgaris as a biofertilizer for maize

Table 7: Mean proximate analysis of Zea mays exposed to the highest CV vulgaris biomass concentration (1.5%)
and the recommended NPK fertilizer dose
Treatments Proximate analysis of maize (%)
Fat Protein Fibre Carbohydrate
C. vulgaris (1.5%) 8.32±0.02a 9.87±0.34a 10.4±0.82a 55.49±0.90a

NPK Fertilizer (100%) 9.99±0.10a 9.74±0.50a 10.65±1.01a 54.2±0.82a

Means having the same letter(s) per column are not significantly different (p>0.05).

DISCUSSION
In the present study, the levels of nitrogen, yield parameters in both dry and rainy seasons. The
phosphorus and potassium in soils treated with C. results obtained by combining C. vulgaris with the
vulgaris and NPK fertilizer were significantly chemical fertilizer at 50 and 100% the
different from the control. This implies that the recommended dose are very encouraging. Thus, at
presence of C. vulgaris in soil is capable of altering 50% the recommended dose of chemical fertilizers
the chemical composition of soil. This could be in combination with 1.5% C. vulgaris, it is possible
related to the over 100% increase in nutrient to reduce the quantity of chemical fertilizers
content recorded in soils treated with the microalga utilized in the cultivation of maize. This is due to
compared to the control. In agreement with our the fact that the growth, biomass and yield values
findings, [28] reported that C. vulgaris interacts obtained with the combination of 50% of the
with bacteria to amend soil characteristics. recommended dose of chemical fertilizer and 1.5%
Furthermore, the high amount of macro and C. vulgaris were similar to those obtained at the
micronutrients present in C. vulgaris [9, 12, 13, 14, recommended dose chemical fertilizer. [17]
29] contributes to their ability to alter soil obtained similar results after growing pea plant in
characteristics. This agrees with the results of [15], soil inoculated with 1.5% cyanobacterium
who found that C. vulgaris increased micronutrients (Oscillatoria angustissima) and 100% chemical
level in soil, which led to increased growth of fertilizer. The presence of algae in soil can
Bermuda grass. Similarly, the increase in Mn, Fe, stimulate synthesis of GA3 [31] and auxin and
Cu and Zn in CV and chemically fertilized soils cytokinin [10] in germinating plants. As previously
with regards to the control could be as a result of stated, the increase in N and P levels in the soil
the increase in the soil fertility, this is in accordance after exposure to C. vulgaris and chemical
with report by [30] that a fertile soil has the ability fertilizers may be responsible for the increased
to supply essential nutrients in adequate amount growth and yield parameters. These nutrients are
and proportion for plant growth and reproduction. important structural and functional elements that
The results obtained showed a significant can limit the extent to which cell division and
increase in growth and yield parameters of Zea enlargement occurs in plants, thereby leading to
mays after exposure to C. vulgaris in the rainy and increased agronomic parameters [32, 33].
dry season months. The changes observed in maize [33]reported that the combination of biofertilizer
growth and yield parameters can be related to and chemical fertilizer leads to large leaf area and
improved nutrient status of soil caused by the leaf number of cowpea, due to high nutrient
presence of C. vulgaris, regardless of the season. availability to the plant. Also, the increase in leaf
Although, [29] worked with dead C. vulgaris area in response to the different fertilization
biomass, the author obtained similar results to those treatments may have led to accumulation of
reported in our study with maize. On the other pigments in leafs [17]. Furthermore, pre-application
hand, [1] reported that soil inoculation with living of C. vulgaris at 1.5% increased agronomic
cells of C. vulgaris enhanced nutrient uptake by parameters compared with the post-application
Lettuce. They reported that this enhancement was which could be as a result of longer time of action
due to the conversion of complex nutrients into prior to planting of the crop. Similar results were
simpler forms, which made them readily available obtained by [29] in maize (Triple-hybrid 310)
for plants and led to increased leaf area, leaf inoculated with dry CV vulgaris who showed that
number and total biomass. Unfortunately, the pre- inoculation of 1.5% CV vulgaris in the soil 15 days
and post-application of only C. vulgaris to maize before planting increased plant height.
gave results that were significantly lower than those The combination chemical fertilizer and C.
we obtained with the NPK chemical fertilizer or a vulgaris had a significant effect on pigment
combination of both. production by Z. mays in both seasons. Pigments
For the first time, our study showed that production and quality are important indicators of
the combination of C. vulgaris with chemical can the physiological state of plants [32]. The
result in significant changes in plant growth and accumulation of these pigments in response to

Nigerian Journal of Scientific Research, 16 (5): 2017; September- October; njsr.abu.edu.ng 629
Sambo et al. (2017); Chlorella vulgaris as a biofertilizer for maize

individual and combined effects of C. vulgaris and growth of competing species [10]. Therefore, it is
chemical fertilizer, may have contributed to the not completely surprising to observe that the
comparable proximate composition of maize seeds combination of C. vulgaris with NPK fertilizer
obtained from the separate exposure to either the tended to have positive effects on the growth of
microalga or NPK fertilizer at 100%. This result is maize plants.
in agreement with that obtained by [34], who found
that the protein content of grains of P. aureus and P. CONCLUSION
mungu exposed to Spirulina were not different from
that of the control. Furthermore, [35] showed that The results of the present study indicate improved
individual or combined exposure to chemical and status of nitrogen, phosphorus and potassium in C.
biological fertilizers sometimes resulted in vulgaris and chemical fertilizer treated soils
significant changes in plant physiology, which compared to control. Pre-application of C. vulgaris
determined the amount of photosynthetic products at a concentration of 1.5% increased crop yield (cob
accumulated. These products served as important length and number of seed rows in maize) and other
raw materials for the biosynthesis of proteins, agronomic parameters, than the post application
carbohydrates and lipids [36], which are transported without chemical fertilizer. The combination of
to stems, roots and seeds. For example, [37] 1.5% C. vulgaris with 50 and 100% chemical
demonstrated that cyanobacterial suspension has fertilizer resulted in high yield and growth
biologically active compounds including plant parameters of Zea mays. The application of CV
growth regulators, which can decrease senescence vulgaris at 1.5% and chemical fertilizer at 50% can
and transpiration and increase the content of leaf provide an alternative way of drastically reducing
chlorophyll. Furthermore, green microalgae have the amount of chemical fertilizer required for maize
also been shown to produce and release bioactive cultivation.
compounds into the environment to regulated

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