Int. J. Aquat. Biol. (2019) 7(2): 95-99

ISSN: 2322-5270; P-ISSN: 2383-0956

Journal homepage: www.ij-aquaticbiology.com
© 2019 Iranian Society of Ichthyology

Original Article
Effect of Chlorella vulgaris as a biofertilizer on germination of tomato and cucumber

seeds

Odgerel Bumandalai*,1Rentsenkhand Tserennadmid

Microbial Synthesis Laboratory, Institute of General and Experimental Biology, Mongolian Academy of Sciences, Mongolia.

Article history:
Received 27 February 2019

Accepted 16 April 2019

Available online 2 5 April 2019

Keywords:
Biofertilizer

Germination

Algae

Crop

Abstract: Although inorganic fertilizers are known to raise environmental and health problems, the
current agricultural practices are heavily dependent on the application of synthetic fertilizers and

pesticides. In this study, we examined the effect of Chlorella vulgaris strain on germination of tomato

and cucumber seeds. Seeds were germinated in culture medium containing algal strain and grown for

3, 6, 9 and 12 days to study its effect on growth parameters. As results, C. vulgaris suspension

increased the seed growth compared to those of the control (sterilized culture medium) of seed

germination. The best treatments were 0.17 and 0.25 g/L of algal suspension for the root and shoot

lengths of tomato and cucumber seeds, respectively.

Introduction

As the global food demand increases, agriculture

sectors have been increasingly using chemical

fertilizer. Inorganic fertilizers are rich in chemical

substances, such as nitrogen, phosphorus and

potassium. The excess uses of chemical fertilizers in

agriculture are costly and also have various harmful

effects on both living organisms and environment

(Santos et al., 2012). For instance, residual chemicals

reach to water bodies through rainwater and cause

eutrophication in water bodies. It can also reduce

water-holding capacity, soil fertility and disparity in

soil nutrients. Moreover, groundwater contamination

could lead to gastric cancer goiter, metabolic disorder,

birth malformations, hypertension and livestock

poisoning (Khandare, 2013; Youssef and Eissa, 2014).

In this regard, organic fertilizers and biofertilizers

have become alternative sources.

Biofertilizers are eco-friendly, cost effective and

renewable source of plant nutrients to supplement and

replace the chemical fertilizers for sustainable

agriculture (Raja, 2013). Biofertilizers contain various

microorganisms that provide all kinds of micro and

macro-elements via nitrogen fixation, phosphate and

*Correspondence: Odgerel Bumandalai DOI: https://doi.org/10.22034/ijab.v7i2.582

E-mail: odgerelb@mas.ac.mn

potassium solubilization or mineralization, release of

plant growth promoting substances, production of

antibiotics and biodegradation of organic matter in the

soil (Goel et al., 1999; Sinha et al. 2010). When

biofertilizers are used continuously for many years,

parental inoculums become sufficient for further

multiplication (Youssef and Eissa 2014), hence they

participate in nutrient cycling and benefit crop

productivity (Singh et al., 2011). Main benefits of

biofertilizers are (1) cheap source of nutrients, (2)

suppliers of microelements, (3) suppliers of organic

matter, (4) counteracting negative impact of chemical

fertilizers, (5) secretion of growth hormone (Gaur,

2010), (6) no adverse effects to ecosystem and (7)

longer shelf life (Sahoo et al., 2014).

The main microorganisms used in biofertilizers are

Azotobacter, Azospirillium, cyanobacteria, Azolla,
phosphate solubilizing microorganisms, mycorrhizae,

Sinorhizobium and plant growth promoting
Rhizobacteria (Hegde et al., 1999; Youssef and Eissa
2014). Algal biomass contains macronutrients as well

as micronutrients, growth regulators, polyamines,

natural enzymes, carbohydrates, proteins, amino

acids, and vitamins implemented for improving

Bumandalai and Tserennadmid / Effect of Chlorella vulgaris as a biofertilizer on tomato and cucumber

vegetative growth (Abd El Moniem and Abd-Allah,

2008; El-Fouly et al., 1992; Mahmoud and Amara,

2000; Shaaban and Mobarak, 2000). In addition, algal

biomass increases the yield due to the presence of

vitamins, auxins and gibberellins. The algal varieties

tested as biofertilizers primarily belong to the branch

of blue-green algae (Cyanophyta) and green algae

(Chlorophyta). It has been shown in many studies that

green algae can (1) add organic matter, (2) synthesize

and liberate amino acids, vitamins and auxins, (3)

reduce oxidizable matter content of the soil, (4)

provide oxygen to the submerged rhizosphere, (5)

improve salinity and buffer the pH, (6) solubilize

phosphate, and (7) increase the fertilizer use efficiency

of crop plants (Faheed and Abd-El Fattah, 2008; Abd

El Moniem and Abd-Allah, 2008; Bileva, 2013;

Dubey and Dubey, 2010; Grzesik and Romanowska-

Duda, 2015; Vig et al., 2012).

The aim of this work was to study the effect of

Chlorella vulgaris strain on germination of tomato and
cucumber seeds and to determine any potential

application of C. vulgaris microalga as a biofertilizer
to improve the yield quality and productivity.

Materials and Methods

Plant material: The experimental plants were seeds of

tomato and cucumber. These seeds were purchased

from retailer store under Ministry of Food,

Agriculture, Light Industry, Mongolia in 2017 and

kept at -4°C under dark condition until the experiment.

Algal culture: Microalga strain was obtained from the

Culture Collection of Microalgae at Institute of

General and Experimental Biology and cultivated

using standard medium 04. The final pH of the

medium was 6.8, after being autoclaved. The culture

was grown with a light intensity of 8 Klux provided

by cool white fluorescent lamps and a temperature of

25±2°C under illumination regime of 8:16 light and

dark cycle for a week. Filtered air was let to bubble in

the culture vessels to provide aeration and agitation.

Effect of culture media after growth of algal strain on

seed germination. Algal suspensions were collected at

3d, 6th, 9th and 12th days and examined for both cell

count and dry biomass yield.

Determination of cell number and biomass content of

alga. Growth of C. vulgaris strain was measured in
terms of cell number and dry weight biomass. Cell

concentrations were counted using a hemocytometer.

Data were given as cell per mL. The determination of

dry biomass yield was performed using Vladimirova’s

method (Sirenko, 1975). The culture suspensions were

mixed well prior to the sampling. 5 mL of samples

were collected in weighing bottles thrice weekly. The

bottles were dried at 105°C oven until the weight of

the bottles become constant. The dry biomass yield

was determined using following formula:

DW (g/L) = (a – b)/Y × 200

Where a is total weight of weighing bottle

containing dried biomass (g), b=weight of the

weighing bottle (g) and Y=sample volume taken (mL).

The data were given as mg/g algae mass.

Treatment of tomato and cucumber seeds. Seeds were

surface sterilized with 30% sodium hypochlorite for 8

min, then rinsed with distilled water several times

before germination. The seeds of tomato and

cucumber were placed in petri dishes containing 3 mL

of sterilized culture medium as a control. 2 ml of algal

suspension was collected after growing the algal strain

for 3, 6, 9 and 12 days, and added to each petri dish

containing tomato and cucumber seeds. Petri dishes

were maintained in thermostat at temperature of

18±2°C under the light regime of 8:16 light and dark

for a week. At the end of the experiment, lengths of

shoots and roots per plant were determined.

Statistical analysis. All experimental analyses were

performed in triplicate and the mean values were

calculated. The data were subjected to analysis of

variance and Student’s t-test and F-test were used to

assess differences between means.

Results

Growth parameters (cell counts and dry biomass

yield) of alga. The growth of algal strain was followed

after 12 days. The cell counts and dry weight were

recorded at 3, 6, 9, and 12 days. As shown in Table 1,

the dry biomass yields were 0.06, 0.12, 0.17 and 0.25

g/L at day 3, 6, 9 and 12, respectively. Meanwhile, the

Int. J. Aquat. Biol. (2019) 7(2): 95-99

cell counts were 26.6, 50.1, 77.2 and 109.8 million

cell/mL in respective algal suspensions.

Seed germination and seedling growth of tomato and

cucumber seeds: The results of growth parameters

obtained for the germination of tomato and cucumber

seeds subjected to culture media after growth of

Chlorella strain for 3, 6, 9, and 12 days treatments are
given in Figures 1 and 2.

The lengths of shoot and root of tomato were

highest at day 9, which were 17.4 and 44.6 mm,

respectively (Fig. 1). As compared to the control, the

growths of roots gradually increased by 29.1, 52.8,

100% at days 3, 6 and 9, respectively. However, the

length of root was 40% shorter than that of control at

day 12. The lengths of shoot at days 6 and 12 were

close to that of control. The lengths of shoots at day 3

and 9 were higher than that of control by 43.3 and

87.9%, respectively.

The lengths of cucumber shoot and root were 2.1

and 2.4 times longer than that of control at day 12,

Table 1. Concentrations of algal suspension used in this experiment.

Days 3 6 9 12

Dry biomass (g/L) 0.06 0.12 0.17 0.25

Cell counts (cells/ml) 26.6*106 50.1*106 77.2*106 109.8*106

Figure 1. Effect of culture medium containing Chlorella grown for 3, 6, 9 and 12 days on growth parameters of seed germination of tomato.

Figure 2. Effect of culture medium containing Chlorella grown for 3, 6, 9 and 12 days on growth parameters of seed germination of cucumber.

Bumandalai and Tserennadmid / Effect of Chlorella vulgaris as a biofertilizer on tomato and cucumber

which were 21.6 and 55 mm, respectively (Fig. 2). The

lengths of shoot were close to that of control at day 3,

6, and 9. The length of root was close to that of control

at day 3. However, the lengths of roots were 41.5 and

80.8% higher than the control at days 6 and 9.

Discussions

Tomato and cucumber are considered as the most

important and common vegetable plants in many

countries. Many studies have been conducted on

tomato to develop bio-stimulants, which can improve

lateral and longitudinal root formation, roots nutrient

uptake, increasing total volume and vigor of the root

system. Among them, Chlorella microalga was also
tested for the tomato bio-stimulant. In our study, the

result showed the significant growth of shoot and root

in tomato plant. The similar results were also observed

in other researchers’ studies. The application of

C. vulgaris was tested on tomato plant, which showed
strong stimulating effect on plant growth while

inhibiting the development of Meloidogyne arenaria
nematode parasite (Choleva et al., 2005). It was also

observed in Garcia-Senin’s (2013) study that the use

of irrigation water with C. pyrenoidosa and Chlorella
sp. cultures favored the production of tomato plants

with special attention in poor soils. Author also

concluded that their results could lead to a lower

environmental impact and a cost-effective tomato crop

production. Moreover, several ocean algae extracts

were tested on tomato seedlings and their results also

showed the enhanced seed germination, plant growth,

and germination rate (Hernández-Herrera et al., 2014;

Shariatmadari et al., 2011). The similar phenomenon

was also observed in cucumber plant. The results

showed the growth of root in cucumber plant was

increasing as the concentration of microalgal

suspension increases. In the study of Abd Elhafiz et al.

(2015), Chlorella sp. cultures were shown to enhance
the germination of cucumber seeds.

Conclusion

It can be concluded that C. vulgaris suspension can
enhance the germinations of tomato and cucumber

seeds. Algal suspensions of 0.17 and 0.25 g/L can

improve the root and shoot lengths of tomato and

cucumber seeds, respectively.

Acknowledgements

We thank Mr. Ghorbanzade, Ghane and Darand for

sampling.

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