Le T. Xa, Nguyen K. Nghia.

Ho Chi Minh City Open University Journal of Science, 10(1), 53-64 53

Microbial diversity of indigenous microorganism communities

from different agri-ecosystems in Soc Trang province, Vietnam
Le Thi Xa1, Nguyen Khoi Nghia2*
1
PhD. student at Biotechnology Research and Development Institute, Can Tho University,
Can Tho City, Vietnam
2
Department of Soil Science, College of Agriculture, Can Tho University, Can Tho City,
Vietnam
*
Corresponding author: nknghia@ctu.edu.vn

ARTICLE INFO ABSTRACT

DOI:10.46223/HCMCOUJS. Indigenous Microorganism (IMO) has great potential for

tech.en.10.1.360.2020 agricultural uses since they have high ability in biodegradation,
nitrogen fixation, phosphate solubilization, plant growth hormone
production as well as bio-control. This study aimed to determine
the presence of some different major groups of microbes in IMO
from different Agri-ecosystem habitats like bacteria, fungi,
actinomyces, Salmonella, Shigella, E. coli, and Coliform. The
presence of bacteria, actinomyces, and fungi of IMO samples was
identified by 27F/1492R, 243F/1378R and ITS1F/ITS4R primers,
respectively. The numbers of bacteria, fungi, and actinomyces
Received: October 1st, 2019 were determined by the plate counting method on TSA, PDA and
Starch media, respectively. The numbers of Salmonella sp. and
Revised: December 24th, 2019
Shigella sp. were determined by the plate counting method on
Accepted: December 26th, 2019
selective Salmonella and Shigella agar (SS agar) after incubation
for 48 hours at 37oC while the density of Coliforms sp. and E.coli
were counted by the Most Probable Number method (MPN). The
results of the study showed that 3 major groups of microbes
including bacteria, fungi, and actinomyces in 14 collected IMO
samples were detected genetically. Moreover, bacterial numbers
were dominated and ranged from 106 to 109cfu/g IMO samples
while the density of fungi and actinomyces were lower and varied
from 105 to 107cfu/g IMO sample. Interestingly, all surveyed IMO
Keywords: samples did not contain any human disease pathogens such as
bacteria, coliforms, Salmonella, Shigella, Coliforms and E. coli. These results imply
indigenous microorganism that collected IMO contains a high diversity of major groups of
communities, Salmonella, microbes and can be used as safe bio-stimulants for clean
Shigella vegetable production.
54 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64

1. Introduction
A concept of Indigenous Microorganism (IMO) was developed by Cho Han Kyu in the
1960s from the Janong Farming Institute, South Korea. IMO cultures contain consortia of
beneficial microorganisms, mainly comprising fungi, bacteria, and actinomyces that are
collected and cultured from soils to use as bio-fertilizer for the plant (Reddy, 2011). Indigenous
microorganism community (IMO) is a group of the innate microbial consortium that inhabits
the soil and the surfaces of all living things. It has potential for biodegradation, bioleaching,
bio-composting, nitrogen fixation, phosphate solubilization, soil fertility improvement and in
the production of plant growth hormones as well as bio-control (Kumar & Gopal, 2015).
Application of IMO in agriculture is a friendly environmental method and helps to enhance
organic matter decomposition, plant nutrition, soil fertility, crop yields and resistance to plant
diseases (Chiemela, Serafin, Ricardo, & Joseph, 2013b). A study by Chiemela, Serafin,
Ricardo, and Joseph (2013a) showed that IMO had strong efficacy on the promotion of
agricultural and plant residues degradation in a compost heap and produced a large number of
soluble micronutrients that are ready to be taken up by plants. In general, it is well-known that
IMO brings many benefits to plants and has been applied broadly in agriculture in many
countries all over the world. IMO-based technology, a great innovation has been applied widely
in many countries in the eastern part of the world. In Vietnam, the application of IMO in organic
coffee farming in Daklak showed that IMOs helped to improve the physical, chemical and
biological properties of the coffee soils. Moreover, IMOs also reduced 25% chemical fertilizer
of the recommended chemical fertilizer formula, but the productivity of this treatment was not
significantly different from the recommended chemical fertilizer treatment (Pham & Dok,
2009). However, deep and scientific knowledge about IMO microorganism composition and
any harmful bacteria residing in IMO have been still lacked and should be scientifically
elucidated. Therefore, this study aimed to determine the presence of some different major
groups of microbes in IMO from different agro-ecosystem habitats like bacteria, fungi,
actinomyces, Salmonella, Shigella, E. coli, and Coliform.
2. Materials and methods
2.1. Materials
Fourteen different IMO were collected from different crop models in Soc Trang
province, Vietnam including bamboo, crop rotation (corn, watermelon, courgette), banana,
shallot, vegetables, rice, watermelon, grassland, maize, lettuce, oranges, grapefruit, guava,
sugarcane by following the method described by Kyu and Koyama (1997) (Figure 1).
 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64 55

Figure 1. The procedure of IMO collection

Sites of sample collection are presented in Table 1. At each sampling site, three plastic
baskets with a size (25 x 15 x 8 cm) were used, corresponding to 3 replicates. Each basket was
filled with 1 kg of steamed rice and covered on the top of the basket with cloth and waist belt.
The baskets were buried underground at each sampling site and covered the top of the baskets
with leaf litters for three days. After three days of incubation, all fermented rice colonized by
indigenous microorganisms was harvested, put into a glass jar and carried to the laboratory.
This source of microorganisms was called IMO1. Collected IMO1 were mixed with brown
sugar paste with a ratio of 1:1 (w/w) until the mixed material became gooey. This mixed
material was stored in the ceramic pot in a cool area and away from direct sunlight for seven
days for another fermentation time. After seven days of fermentation, this source of
microorganisms was called IMO2 and the IMO2 was kept in the refrigerator at 4oC for further
studies (Le, Nguyen, & Nguyen, 2018).
Table 1
The located of nineteen collected IMO samples in Soc Trang province

Origin of IMO
Code Located in Soc Trang province
(Farming system)
TP-1 Bamboo Phu Tam commune, Chau Thanh district
MQ-2 Crop rotation 5 Ward, Soc Trang city
CP-3 Banana 7 Ward, Soc Trang city
HV-4 Shallot Vinh Phuoc commune, Vinh Chau district
RP-5 Lettuce 3 Ward, Soc Trang city
LM-6 Rice My Xuyen town, My Xuyen district
DL-7 Watermelon Truong Khanh commune, Long Phu district
56 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64

Origin of IMO
Code Located in Soc Trang province
(Farming system)
CL-8 Grassland Truong Khanh commune, Long Phu district
BT-9 Maize Thanh Tri commune, Thanh Tri district
RM-10 Vegetables Thanh Quoi commune, My Xuyen district
CK-11 Oranges Xuan Hoa commune, Ke Sach district
BK-12 Grapefruit Xuan Hoa commune, Ke Sach district
OK-13 Guava Xuan Hoa commune, Ke Sach district
MC-14 Sugarcane Đai An II commune, Cu Lao Dung district
Source: The researcher’s data analysis

2.2. Determining the presence of bacteria, fungi, and actinomyces in different IMO2
Firstly, the total DNA of each IMO2 was extracted by MO BIO kit (MOBIO
Laboratories, QIAGEN Company, Valencia, CA), then used 27F/1492R primers for PCR
reaction to amplify 1500bp sequences of 16S rARN of bacteria (Lane, 1991). The nucleotide
sequence of 27F primer includes 5’AGA GTT TGA TCC TGG CTC AG3’ and the nucleotide
sequence of 1492R contains 5’GGT TAC CTT GTT ACG ACT T3’. All PCR amplifications
were carried out in 20μL reactions containing 10μL Green mix (2X), 1μL 27F primer (10μM),
1μL 1492R primer (10μM), 2μL pure DNA and 6.0μL deionized water. All reactions were run
on the GeneAmp PCR System 9700. Samples were amplified using the following parameters:
5-min initial denaturation of DNA at 95°C, followed by 30 cycles of 1-minute denaturation at
94°C, 1-minute primer annealing 53oC, and 90-minute extension at 72°C. Amplification was
completed by a final extension step at 72°C for 7 minutes. In addition, ITS1/ITS4 primers were
used for PCR reaction to amplify 675bp sequences of the ITS gene region of fungi. Fungal
DNA amplification began with an initial denaturation at 95°C for 5 min, 30 cycles of 96°C for
1min, 63°C for 1min, 72°C for 1min and followed by the step of 72°C for 7min (Tao, Liu,
Hyde, Lui, & Yu, 2008). The nucleotide sequence of ITS1 and ITS4 primers contains 5’-
CTTGGTCATTTAGAGGAAGTAA-3’ and 5’-TCCTCCGCTTATTGATATGC-3’,
respectively. Finally, 243F/1378R primers were used for PCR reaction to amplify 1175bp
sequences of 16S rARN actinomyces (Heuer, Krsek, Baker, Smalla, & Wellington, 1997). The
actinomyces DNA started with an initial denaturation step of 95°C for 5min before 30 cycles
of 94°C for 1min, 63°C for 1min, 72°C for 2min and then a final step of 72°C for 10min for the
last cycle. To visualize the PCR products, 5µL of the reactions were loaded into 1.5% of agarose
gel, 5µL of 100bp DNA ladder was also loaded into a gel as a molecular weight biomarker.
Gels were run for 30 minutes at 150 volts and 500 milliamps and then visualized and
photographed by UV light from Gel Logic 1500 (Kodak) to find target sequences with the size
of 1465bp for bacteria, 675bp for fungi and 1175bp for actinomyces, respectively.
 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64 57

2.3. Cell counting of bacteria, fungi, and actinomyces in collected IMO2

An aliquot of 10 grams of each IMO2 was put into a 250mL glass bottle containing
90mL sterilized distilled water on a shaker at a speed of 150 rpm for an hour and left stand for
5 minutes after shaking. A series dilution with a factor of 10 was prepared. An aliquot of 50µL
of each dilution was spread on PDA, TSA, and Starch agar plates as media for bacteria, fungi,
and actinomyces, respectively. Each dilution of each medium was repeated triply. The
composition of PDA medium (g/L) contained 200g potato infusion, 20g dextrose, 20g agar for
1 L distilled water. The composition of TSA medium (g/L) comprised 17g tryptone, 3g soya
peptone, 5g sodium chloride, 2.5g dipotassium hydrogen phosphate, 2.5g dextrose. The
composition of 1liter Starch medium containing 20 g Starch, 1.15g K2HPO4, 1.5gMgSO4, 1g
NaCl, 2g (NH4)2SO4, CaCO3 and 20g agar. In addition to (500 IU/L) streptomycin
antimicrobial compound, TSA supplemented with 500 IU/L nystatin antifungal compound for
bacteria and Starch medium supplemented with K2Cr2O7 0.4%for actinomyces. Samples were
placed in incubators at 30oC for 3-7 days. Finally, the number of microbes developed on each
agar medium was counted to calculate the numbers of colony-forming units.
2.4. Determination of the presence of Coliforms, E. coli, Salmonella, Shigella spp. in
collected IMO2
Salmonella and Shigella numbers were determined by counting their colonies on
selective media for only Salmonella and Shigella (SS agar) after incubation for 48 hours at 37oC
(Taylor & Harris, 1965). An aliquot of 100µL IMO2 solution (prepared in section 2.3) was
transferred and spread onto the SS medium agar. Three replications were repeated for each
dilution. All agar plates were placed in an incubator at 37°C and the presence of Salmonella
and Shigella colonies after 48 hours of incubation was examined.
Coliform numbers were determined by transferring 0.4mL of each dilution
concentration (in 2.3) into a 20mL test tube containing 3.6mL of Lauryl Sulphate Broth medium
(LSB) and an upside-down Durham tube (5 replications for each diluted concentration). The
composition of Lauryl Sulphate Broth medium (g/L) included 20g Tryptose, 5g lactose, 5g
NaCl, 2.75g KH2PO4, 2.75g K2HPO4, and 0.1g Sodium lauryl sulfate. The tubes were
incubated at 34.5oC, for 48 hours and then observed and recorded the presence of air bubbles
in the sample. The sample was identified as negative for Coliform when air bubbles did not
exist in the liquid medium of the test tubes and the samples were considered positive for
Coliform when air bubbles appeared in the liquid medium of the test tubes. To quantify the
numbers of E.coli in the sample, transferred 0.4mL liquid media of test tubes showing positive
Coliform into a new test tube containing 3.6mL of Escherichia coli broth (EC broth) and upside-
down Durham tube with no air bubbles. The composition of EC broth medium containing 20g
tryptose, 1.5g of Bile salts No.3, 5g lactose, 1.5g KH2PO4, 4g K2HPO4, 5g NaCl, pH 6.8 in 1
litter. The test tubes were incubated in an incubator for 24-hour at 44.5°C. The sample was
identified as negative for E.coli when air bubbles did not exist in the liquid medium of the test
tubes and the samples were considered positive for E.coli when air bubbles appeared in the
liquid medium of the test tubes. The density of Coliforms sp. and E.coli were counted by the
Most Probable Number method (MPN).
58 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64

2.5. Data analysis

The data were analyzed by ANOVA and compared by the DUNCAN test with
MINITAB version 16 software.
3. Results and discussion
3.1. The presence of bacteria, fungi, and actinomyces in collected IMO2
The results of a polymerase chain reaction for testing the presence of three main groups
of microorganisms including bacteria, fungi, and actinomyces are shown in Figure 2 indicating
that all three main groups of microorganisms were detected in all collected IMO2 samples. For
bacteria, the 27F/1492R primers amplified successfully a fragmented DNA with a 1475bp
length of bacteria’s 16S rARN gene from DNA of all collected IMO2 (Figure 2A). This result
reveals that there was a presence of bacterial community in all collected IMO2 samples from
the different farming ecosystems, the presence of bacteria and the clear visual band with
different intensities of each well in agarose gel electrophoresis of Figure 2 indicated for the
different population of the bacterial community of IMO2. Similarly, all collected IMO2 were
recorded to have the size of target sequences (650bp) when running PCR by ITS1F/ITS4R
primers for fungi (Figure 2B). Therefore, the fungi community was also presented in all 14
collected IMO2 samples. Moreover, special primers for actinomyces, 243F/1378R also
successfully amplified target sequences (1150bp) of the 16S rARN gene for actinomyces in
IMO2 (Figure 2C). This proved that in all IMO2 samples collected there was the presence of
actinomyces, although it is hard to know which species and genera group of actinomyces. In
general, the results of this study suggest that all IMO2 collected from different ecosystem
habitats have a very high microbial diversity including bacteria, fungi, and actinomyces as three
main groups of microbes in IMO2 samples.
This result was consistent with the study of Kyu and Koyama (1997), Kalsom and Sariah
(2006) and Reddy (2011). These studies indicated that indigenous microorganisms composition
containing bacteria, fungi, actinomyces, nematodes, protozoa, ... and the activities of these
microbial groups would help to increase biodegradation, nitrogen fixation, phosphorus
dissolution and synthesis of plant growth promotion, soil fertility recovery and improvement
and organic matter decomposition in the soil. Consequently, crop yields will be improved,
reducing pathogenic microorganisms and increasing plant defense.
 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64 59

Figure 2. PCR products of specific sequences for bacteria (A), fungi (B) and actinomyces (C)
by special primer amplification for 14 collected IMO
*
Note: Lane 1: 100bp standard ladder; lane 2: IMO from bamboo; lane 3: IMO from crop rotation; lane 4: IMO
from banana; lane 5: IMO from shallot; lane 6: IMO from vegetables; lane 7: IMO from rice; lane 8: IMO from
watermelon; lane 9: IMO from grassland; lane 10: IMO from maize; lane 11: IMO from lettuce; lane 12: IMO
from oranges; lane 13: IMO from grapefruit; lane 14: IMO from guava; lane 15: IMO from sugarcane

3.2. The numbers of bacteria, fungi, and actinomyces in collected IMO2

The numbers of bacteria, fungi, and actinomyces in fourteen different IMO2 are
presented in Table 2. The numbers of 3 main groups of microorganisms within each indigenous
microorganism were different and between collected IMO from different soil ecosystems were
also different. In general, in each IMO, the bacterial population was significantly higher than
the other two groups, and the different microbial composition between IMO was accumulated
from different agro-ecosystems.
3.2.1. Bacteria
It can be seen from Table 2 that the number of bacteria in IMO2 obtained from different
habitats varied significantly when compared with each other. Bacterial numbers in the IMO
ranged from 1.36 x 107 to 2.13 x 109cfu/g IMO in which IMO collected from guava soil has the
highest numbers of bacteria (2.13 x 109cfu/g IMO) and was significantly higher than bacterial
60 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64

numbers of the other remaining IMO. The IMO collected from maize, vegetables, and
sugarcane cultivated fields together shared second place with the same numbers of bacteria for
all three treatments (32 x 107cfu/g IMO). The lowest number of bacteria was found in IMO
collected from crop rotation system, banana, and shallot cultivated fields with several 3.70 x
107, 2.15 x 107 and 4.75 x 107cfu/g IMO, respectively while the other remaining IMO had higher
numbers of bacteria and ranged from 8.45 x 107 to 28.40 x 107cfu/g IMO.
Table 2
The microbial density of fifteen indigenous microorganism communities

Origin of Microbial density

samples
Number Bacteria Fungus Actinomyces
(Farming
system) (107cfu/g IMO) (106cfu/g IMO) (105cfu/g IMO)

1 Bamboo 13.30bcd 143.5a 3.67d

2 Crop rotation 3.70cd 20.50cd 2.80d
3 Banana 2.15d 91.50b 1.93d
4 Shallot 4.75cd 27.00cd 2.93d
5 Lettuce 13.60bcd 2.05d 51.33bc
6 Rice 13.15bcd 51.50bc 113.3a
7 Watermelon 24.90bcd 140.0a 118.0a
8 Grassland 21.05bcd 2.05d 61.33b
9 Maize 32.50b 3.00d 30.00cd
10 Vegetables 32.35b 3.55d 24.67cd
11 Oranges 28.40bc 2.65d 2.60d
12 Grapefruit 8.45bcd 24.00cd 2.60d
13 Guava 212.50a 93.50b 3.27d
14 Sugarcane 32.45b 32.50cd 18.00d
15 Mix 11.75bcd 30.50cd 24.00cd
*Note: Values in the same column having the same letters are not a significant difference at 5% level (p<0.05)
Source: The researcher’s data analysis

3.2.2. Fungi
The results presented in Table 2 show that the fungal density in all IMO was lower than
the bacterial density and among different IMO, the fungal density was significantly different
from each other. The fungi’s numbers of IMO collected from bamboo plantation was highest
with the number of 143 x 107cfu/g IMO, and next, the number of fungi in IMO collected from
 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64 61

bananas and guava farms were 91.5 x 106 and 93.5 x 106cfu/g IMO and significantly higher
than those of the remaining IMO. Followed by IMO collected from the rice field with a fungal
number of 51.5 x 106cfu/g IMO. In contrast, the other five IMOs collected from lettuce,
grassland, maize, vegetables, and oranges farms had the lowest density of fungi with a variation
between 2.05 x 106 and 3.55 x 106cfu/g IMO while the remaining IMO had fungal numbers
ranged from 20.5 x 106 to 30.5 x 106cfu/g IMO.
3.2.3. Actinomyces
The numbers of actinomyces in fifteen different IMO are presented in Table 2. The
results illustrated that the numbers of actinomyces were significantly lower than that of bacteria
and fungi and between IMO collected from different soil ecosystems; there was also a strongly
significant difference among each other in terms of bacterial numbers. Particularly, two IMO
collected from rice and watermelon farms had the highest numbers of actinomyces with several
113.0 x 105 and 118.0 x 105cfu/g IMO, respectively. While the lowest density of actinomyces
was found with a range between 1.93 x 105 and 3.67 x 105cfu/g IMO in IMO from bamboo,
crop rotation, banana, shallot, oranges, grapefruit, and guava farms. Other remained IMO had
numbers of actinomyces varied between 18.0 x 105 and 61.33 x 105cfu/g IMO.
In a comparison with other previous studies, Chiemela et al. (2013a) indicated that in
the bamboo forest IMO there was a richness of bacterial population with a number at 2.8 x
109cfu/g IMO, followed by the fungi at 4.2 x 104cfu/g IMO. Also, according to this study, the
growth of microorganisms in IMO depended on various factors such as origin, temperature, pH,
incubation period, carbon, moisture, etc. Similarly, Abu-Bakar and Ibrahim (2013) evaluated
the numbers of bacteria in composts and showed that the number of bacteria was increased
very, from 3.1 x 107 cfu/g to 3.1 x 108cfu/g while the numbers of fungi were decreased
drastically from 1.8 x 108cfu/g to 1.6 x 106cfu/g after day 10 incubation days. The numbers of
actinomycetes were slightly dropped during the composting process. However, it is shown that
the total of the actinomycetes population remained high throughout the entire composting
process until the curing phase.
This result revealed that there was a presence of high numbers of three main microbial
groups in all fourteen studied IMO and this result implies that almost all IMO could be
considered as a good source of beneficial microbes for soil improvement as well as plant growth
promotion. According to Reddy (2011), indigenous microbes are good sources of microbes for
farming because they are very powerful and effective. They can survive better under the
extreme climatic conditions of the local environment than under artificial cultures and
environments. Since they have resided and already adapted to the local conditions, they are
considered to be the best survival source of microbes for soil and plant improvement effectively.
3.3. Coliforms, E. coli, Salmonella, Shigella spp. in the collected IMO2
The results of the experiment showed that there wasn’t any presence of Salmonella and
Shigella in all fifteen collected IMO2 samples. Thus, no Salmonella and Shigella colonies were
detected in SS agar media (Figure 3). It could be that the temperature and pH of the fermentation
process to create IMO samples limited the survival of these two microorganisms in IMO.
62 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64

Figure 3. SS agar medium after 48 hours of incubation

Similarly, for Coliform and E.coli, the results of the survey experiment showed that
there was no presence of Coliform and E.coli in all fifteen collected IMO samples since all the
tested samples did not have any turbidity and bubble in Durham tubes in the samples (Figure
4). Therefore, it can be concluded that Coliform and E.coli don’t exist in our 14 collected IMO
samples. This could be explained by the fact that the environmental conditions like temperature,
pH of samples and other factors during the fermentation process were not good and suitable for
these groups of bacteria to survive and develop.

Figure 4. The test tube containing LSB medium with no air bubbles in Durham tubes both
before and after 48 hours of incubation
It is very interesting that although bacteria are detected with a high density in each IMO,
there are not any harmful bacteria in it. According to Molina et al. (2015), public health
protection requires the prompt evaluation of microorganisms in drinking water, raw and
processed foods as well as bio-fertilizer to prevent outbreaks of microbial contamination.
Therefore, the evaluation of harmful bacteria such as Salmonella, Shigella, Coliforms,
 Le T. Xa, Nguyen K. Nghia. Ho Chi Minh City Open University Journal of Science, 10(1), 53-64 63

particularly Escherichia coli cause sickness for the human is necessary. In short, all surveyed
IMO samples did not have any contamination of human disease pathogens like Salmonella,
Shigella, Coliforms, and E.coli. These results imply that collected IMO contains the high
diversity of major groups of microbes and can be used as safe bio-stimulants for clean vegetable
production.
4. Conclusion
A big community of bacteria, fungi, and actinomyces were detected in all fourteen
collected IMO samples from many diverse origins of habitats by a tool of biological molecular
technology. The largest number of microbes was recorded for bacteria with a range between
1.36 x 107 and 2.13 x 109cfu/g IMO sample whereas the numbers of fungi and actinomyces
varied from 2.05 x 105 to 1.40 x 107 and 1.80 x 105 to 1.18 x 107cfu/g IMO sample, respectively.
Especially, all surveyed IMO samples have not been found to have any contamination of human
disease pathogens such as Salmonella, Shigella, Coliforms, and E.coli. These high diversities
of all collected IMO can be exploited for enhancing soil fertility and plant growth. Moreover,
using IMO for plants as a bio-fertilizer source is safe for human health. IMO is a great potential
source of beneficial microbes that can be used for the isolation of beneficial microbes for bio-
fertilizers to enhance soil health, quality and fertility and for plant growth promotion and
sustainable and clean agricultural development. It is obvious to accept the fact that the study on
the functions of the IOM for agricultural application has still lacked and many outstanding and
good results in this research field are still waiting.

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