ASSESSMENT OF METABOLITES FROM SOIL FUNGI ISOLATED FROM

FACULTY OF PHARMACY BAYERO UNIVERSITY KANO OLD CAMPUS

BY

HAUWA ABDULLAHI GADANYA

UG18BIO1155

SUPERVISED BY

DR AFIYA HAMISU

MARCH, 2024
1
 Chapter One

Introduction

Plants are photoautotrophic organism that involved the water vapor, sunlight, chlorophyll

pigments in mesophyll cells and obtain inorganic nutrient from soil mineral to synthesize

their own organic molecules (Abbas, 1996).

Plants convert the inorganic to organic compounds through the process of photosynthesis.

Photosynthesis means the synthesis of molecules using light, which utilize the complex

carbon molecules. In photosynthesis, energy from the sun is captured and transformed from

light energy into biochemical energy, produces the sugar molecules or glucose and oxygen.

These glucose molecules are involved in the biosynthesis of amino acids for protein, fatty

acids for lipids, and other cell constituents (Anderson et el., 1995).

Photosynthesis takes place in green organism organs which contain the mesophyll cells and

chlorophyll pigment. Plant absorbs the water (H2O) from soil and carbon dioxide (CO2)

present in the air, with the electron molecules emitted from the sunlight. The molecular

binding of water and carbon dioxide will produce the carbohydrate single molecules

(C6H12O6, glucose) in mesophyll cells, and then releases the oxygen gas (O2) that diffuse

out into the atmosphere (Anderson et el., 1995).

These complex macromolecules will be break down into energy and simple inorganic

molecules again, through the process called cellular respiration. Cellular respiration converts

oxygen and glucose into water and carbon dioxide, it takes place in mitochondrial organelles.

Photosynthesis and cellular respiration are responsible for the exchange of oxygen and carbon

dioxide between the living organisms, and these processes are part of plant metabolisms.

Metabolism defined as the essential biochemical processes that takes place in living

organism, correlated with the enzymatic process, require energy to occur and produce macro

molecules in plant storage cells (Strobel et el., 2002).

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Fungi (singular fungus) can be defined as nucleated, achlorophyllous, osmotrophic, spore-

bearing organisms that typically reproduce both sexually and asexually and who’s usually

branching, filamentous bodies are surrounded by cell walls composed of cellulose, chitin or

both. The science or study of fungi is called mycology (mykes = mushroom), and originated

as the study of macroscopic organisms such as mushrooms, puffballs and bracket fungi.

(Abebe et al., 1993).

These organisms were often collected as food items, but the ingestion of toxic fungi

sometimes resulted in fatality and sometimes caused peculiar neurological reactions "they did

funn

y things to the head". It was important that people collecting fungi knew something about

them. Fungi are classified within the domain Eukarya, and according to the Whittaker five-

kingdom system are categorized within the kingdom Fungi (Myceteae); although other

references divide them between the kingdoms Chromista and Eumycotina (Lorthoraly et el.,

2011)

Fungi are eukaryotic organisms with true nuclei surrounded by nuclear envelopes. They are

equipped with a variety of organelles, but unlike plants do not have chloroplasts. The term

achlorophyllous means without chlorophyll (a = without, chlorophyll = green, light- sensitive

pigment). Green pigments are commonly associated with light-trapping ability in green

plants, algae, cyanobacteria and other phototrophic organisms, but fungi do not have this

ability. (Al-Doory, 1984)

1.1 Fungal Metabolites

Fungal metabolites are the diverse array of chemical compounds produced by fungi. These

metabolites play important roles in fungal biology and can have various effects on other

organisms, including humans. Fungi produce a wide range of metabolites with diverse

chemical structures and biological activities.

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Some fungal metabolites have beneficial effects, such as antibiotics, which are produced by

certain fungi to inhibit the growth of bacteria. These antibiotics have been widely used in

medicine to treat bacterial infections. Other fungal metabolites have been studied for their

potential pharmaceutical applications, including antifungal, anticancer, and

immunosuppressive properties.

On the other hand, some fungal metabolites can have detrimental effects on human health.

Certain fungi produce mycotoxins, which are toxic compounds that can contaminate food and

cause various health problems when consumed. Mycotoxins are a concern in food safety, and

their presence in crops and food products is closely monitored to ensure public health.

1.2 Statement of the Problem

Fungi are of great interest due to their ability to produce a wide variety of bioactive

metabolites with potential pharmaceutical and industrial applications. However, the

assessment of these metabolites in fungi is often challenging due to the complex nature of

their metabolic pathways and the large number of metabolites that they can produce. There is

a need for comprehensive and reliable methods for assessing the metabolites produced by

fungi in order to fully explore their potential uses. Current methods for assessing metabolites

in fungi often involve a combination of analytical techniques such as chromatography, mass

spectrometry, and nuclear magnetic resonance spectroscopy. However, these methods can be

time-consuming, costly, and require specialized equipment and expertise. Additionally, the

identification and quantification of specific metabolites can be difficult due to the complexity

of fungal extracts and the presence of numerous co-eluting compounds.

However, the lack of standardized methods for metabolite assessment in fungi makes it

difficult to compare results between different studies and to fully understand the diversity and

abundance of metabolites produced by different fungal species. There is a need for improved

methods for the assessment of metabolites in fungi that are comprehensive, reliable, and user-

friendly. These methods should enable the identification and quantification of a wide range of
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metabolites, as well as facilitate comparative studies between different fungal species. The

development of such methods would greatly advance our understanding of fungal metabolism

and improve the potential for the discovery of novel bioactive compounds.

1.3 Aim and Objectives

1.3.1 Aim

The aim of this project is to assess metabolites in Fungi isolated from Faculty of Pharmacy

Bayero University Kano Old side.

1.3.2 Objectives

i. To isolate purify and identify fungi from the soil

ii. Extract and identify the metabolites from the crude extracts

1.4 Justification

The need to carry out the study is important for understanding the ecology and biodiversity of

Fungal communities, Fungi produce a diverse range of metabolites in response to changes in

environmental conditions. By examining the metabolites produced by different fungal species

in the faculty of pharmacy in Bayero University Kano, the study can gain insights into the

ecological role and interactions of different fungi in the environment (Barron, 1983).

1.5 Definition of Terms

i. Fungi: Fungi are eukaryotic organisms that include microorganisms such as yeasts,

molds, and mushrooms. These organisms are classified under kingdom fungi.

ii. Heterotroph: An organism that cannot make its own food and must obtain nutrients

from other organic sources.

iii. Hyphae: Branching filaments of a fungus.

iv. Mycelium: A network of hyphae.

v. Yeast: Single-celled fungi.

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6
 Chapter Two

Literature Review

2.1 Fungi

Fungi are eukaryotic organisms that include microorganisms such as yeasts, moulds and

mushrooms. These organisms are classified under kingdom fungi. The organisms found in

Kingdom fungi contain a cell wall and are omnipresent.

Fungi or commonly called fungi are microorganisms that have heterotrophic characteristics,

do not contain chlorophyll, are multicellular, produce spores, reproduce sexually and

asexually, generally microscopic, eukaryotic, non-motile, generally saprobes, absorbent,

secrete enzymes to break down organic compounds and store food in the form of glycogen

(Corliss, 2012).

The fungus has a thread-like structure in the form of a tube (cylindrical) called hyphae. There

are hyphae that have a partition (septate) and not insulated (senosit). The collection of hyphae

is called mycelium.

Fungi are usually classified in four divisions: The Chytridiomycota (chytrids), Zygomycota

(bread molds), Ascomycota (yeasts and sac fungi), and the Basidiomycota (club fungi) and

the recently described Phylum Glomeromycota. An older classification scheme grouped fungi

that strictly use asexual reproduction into Deuteromycota, a group that is no longer in use .

Placement into a division is based on the way in which the fungus reproduces sexually. The

shape and internal structure of the sporangia, which produce the spores, are the most useful

character for identifying these various major groups (Corliss, 2012).

2.1.1 Chytridiomycota: The Chytrids

The only class in the Phylum Chytridiomycota is the Chytridiomycetes. The chytrids are the

simplest and most primitive Eumycota, or true fungi. The evolutionary record shows that the

first recognizable chytrids appeared during the late pre-Cambrian period, more than

500 million years ago. Like all fungi, chytrids have chitin in their cell walls, but one group of
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chytrids has both cellulose and chitin in the cell wall. Most chytrids are unicellular; a few

form multicellular organisms and hyphae, which have no septa between cells (coenocytic).

They produce gametes and diploid zoospores that swim with the help of a single

flagellum.The ecological habitat and cell structure of chytrids have much in common with

protists. Chytrids usually live in aquatic environments, although some species live on land.

Some species thrive as parasites on plants, insects, or amphibians, while others are saprobes.

The chytrid species Allomyces is well characterized as an experimental organism. Its

reproductive cycle includes both asexual and sexual phases. Allomyces produces diploid or

haploid flagellated zoospores in a sporangium (Corliss, 2012).

Fig 2.1: Chytridiomycota

Source: MidgleyDJ at en.wikipedia

2.1.2 Zygomycota: The Conjugaed Fungi

The zygomycetes are a relatively small group of fungi belonging to the Phylum Zygomycota.

They include the familiar bread mold, Rhizopus stolonifer, which rapidly propagates on the

surfaces of breads, fruits, and vegetables. Most species are saprobes, living off decaying

organic material; a few are parasites, particularly of insects. Zygomycetes play a considerable

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commercial role. The metabolic products of other species of Rhizopus are intermediates in

the synthesis of semi-synthetic steroid hormones (Corliss, 2012).

Zygomycetes have a thallus of coenocytic hyphae in which the nuclei are haploid when the

organism is in the vegetative stage. The fungi usually reproduce asexually by producing

sporangiospores. The black tips of bread mold are the swollen sporangia packed with black

spores. When spores land on a suitable substrate, they germinate and produce a new

mycelium. Sexual reproduction starts when conditions become unfavorable. Two opposing

mating strains (type + and type –) must be in close proximity for gametangia from the hyphae

to be produced and fuse, leading to karyogamy. The developing diploid zygospores have

thick coats that protect them from desiccation and other hazards. They may remain dormant

until environmental conditions are favorable. When the zygospore germinates, it undergoes

meiosis and produces haploid spores, which will, in turn, grow into a new organism. This

form of sexual reproduction in fungi is called conjugation (although it differs markedly from

conjugation in bacteria and protists), giving rise to the name “conjugated fungi”.

Zygomycetes have asexual and asexual life cycles. In the sexual life cycle, plus and minus

mating types conjugate to form a zygosporangium (Corliss, 2012).

Fig 2.2: Zygomycete Life Cycle

Source: pressbooks-dev.oer.hawaii.edu
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Sporangia grow at the end of stalks, which appear as (a) white fuzz seen on this bread

mold, Rhizopus stolonifer. The (b) tips of bread mold are the spore-containing sporangia.

Fig 2.3: Sporangia Grow

Source: pressbooks-dev.oer.hawaii.edu

2.1.3 Ascomycota: The Sac Fungi

The majority of known fungi belong to the Phylum Ascomycota, which is characterized by

the formation of an ascus (plural, asci), a sac-like structure that contains haploid ascospores.

Many ascomycetes are of commercial importance. Some play a beneficial role, such as the

yeasts used in baking, brewing, and wine fermentation, plus truffles and morels, which are

held as gourmet delicacies. Aspergillus oryzae is used in the fermentation of rice to produce

sake. Other ascomycetes parasitize plants and animals, including humans. For example,

fungal pneumonia poses a significant threat to AIDS patients who have a compromised

immune system. Ascomycetes not only infest and destroy crops directly; they also produce

poisonous secondary metabolites that make crops unfit for consumption. Filamentous

ascomycetes produce hyphae divided by perforated septa, allowing streaming of cytoplasm

from one cell to the other. Conidia and asci, which are used respectively for asexual and

sexual reproductions, are usually separated from the vegetative hyphae by blocked (non-

perforated) septa (Corliss, 2012).

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Asexual reproduction is frequent and involves the production of conidiophores that release

haploid conidiospores. Sexual reproduction starts with the development of special hyphae

from either one of two types of mating strains. The “male” strain produces an antheridium

and the “female” strain develops an ascogonium. At fertilization, the antheridium and the

ascogonium combine in plasmogamy without nuclear fusion. Special ascogenous hyphae

arise, in which pairs of nuclei migrate: one from the “male” strain and one from the “female”

strain. In each ascus, two or more haploid ascospores fuse their nuclei in karyogamy. During

sexual reproduction, thousands of asci fill a fruiting body called the ascocarp. The diploid

nucleus gives rise to haploid nuclei by meiosis. The ascospores are then released, germinate,

and form hyphae that are disseminated in the environment and start new mycelia (Corliss,

2012).

Fig 2.4: Ascomycete Life Cycle

Source: pressbooks-dev.oer.hawaii.edu

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The bright field light micrograph shows ascospores being released from asci in the

fungus Talaromyces flavus var. flavus.

Fig 2.5: Ascospore

Source: pressbooks-dev.oer.hawaii.edu

2.1.4 Basidiomycota: The Club Fungi

The fungi in the Phylum Basidiomycota are easily recognizable under a light microscope by

their club-shaped fruiting bodies called basidia (singular, basidium), which are the swollen

terminal cell of a hypha. The basidia, which are the reproductive organs of these fungi, are

often contained within the familiar mushroom, commonly seen in fields after rain, on the

supermarket shelves, and growing on your lawn. These mushroom-producing basidiomyces

are sometimes referred to as “gill fungi” because of the presence of gill-like structures on the

underside of the cap. The “gills” are actually compacted hyphae on which the basidia are

borne.

This group also includes shelf fungus, which cling to the bark of trees like small shelves. In

addition, the basidiomycota includes smuts and rusts, which are important plant pathogens;

toadstools, and shelf fungi stacked on tree trunks. Most edible fungi belong to the Phylum

Basidiomycota; however, some basidiomycetes produce deadly toxins. For

example, Cryptococcus neoformans causes severe respiratory illness(Corliss, 2012).

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The fruiting bodies of a basidiomycete form a ring in a meadow, commonly called “fairy

ring.” The best-known fairy ring fungus has the scientific name Marasmius oreades. The

body of this fungus, its mycelium, is underground and grows outward in a circle. As it grows,

the mycelium depletes the soil of nitrogen, causing the mycelia to grow away from the center

and leading to the “fairy ring” of fruiting bodies where there is adequate soil nitrogen.

The lifecycle of basidiomycetes includes alternation of generations. Spores are generally

produced through sexual reproduction, rather than asexual reproduction. The club-shaped

basidium carries spores called basidiospores. In the basidium, nuclei of two different mating

strains fuse (karyogamy), giving rise to a diploid zygote that then undergoes meiosis. The

haploid nuclei migrate into basidiospores, which germinate and generate monokaryotic

hyphae. The mycelium that results is called a primary mycelium. Mycelia of different mating

strains can combine and produce a secondary mycelium that contains haploid nuclei of two

different mating strains. This is the dikaryotic stage of the basidiomyces lifecyle and and it is

the dominant stage. Eventually, the secondary mycelium generates a basidiocarp, which is a

fruiting body that protrudes from the ground—this is what we think of as a mushroom. The

basidiocarp bears the developing basidia on the gills under its cap (Corliss, 2012).

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 Fig 2.6: Basidiomycete Life Cycle

Source: pressbooks-dev.oer.hawaii.edu

2.1.5 Glomeromycota

The Glomeromycota is a newly established phylum which comprises about 230 species that

all live in close association with the roots of trees. Fossil records indicate that trees and their

root symbionts share a long evolutionary history. It appears that all members of this family

form arbuscular mycorrhizae: the hyphae interact with the root cells forming a mutually

beneficial association where the plants supply the carbon source and energy in the form of

carbohydrates to the fungus, and the fungus supplies essential minerals from the soil to the

plant.

The glomeromycetes do not reproduce sexually and do not survive without the presence of

plant roots. Although they have coenocytic hyphae like the zygomycetes, they do not form

zygospores. DNA analysis shows that all glomeromycetes probably descended from a

common ancestor, making them a monophyletic lineage (Corliss, 2012).

2.2 Metabolites

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Metabolites are the intermediate products produced during metabolism, catalyzed by various

enzymes that occur naturally within cells. Eg., antibiotics, and pigments. The term

metabolites are usually used for small molecules. The various functions of metabolites

include; fuel, structure, signaling, catalytic activity, defense and interactions with other

organisms. The metabolites are produced by plants, humans and microbes (Corliss et el.,

1994).

Plant Metabolites: Plant metabolites are of two types;

i. Primary Metabolites

ii. Secondary Metabolites

2.2.1 Primary Metabolites

Primary metabolites are involved in growth, development, and reproduction of the organism.

The primary metabolite is typically a key component in maintaining normal physiological

processes; thus, it is often referred to as a central metabolite. Primary metabolites are

typically formed during the growth phase as a result of energy metabolism, and are deemed

essential for proper growth. Examples of primary metabolites include alcohols such as

ethanol, lactic acid, and certain amino acids. Within the field of industrial microbiology,

alcohol is one of the most common primary metabolites used for large-scale production.

Specifically, alcohol is used for processes involving fermentation which produce products

like beer and wine. Additionally, primary metabolites such as amino acids– including L-

glutamate and L-lysine, which are commonly used as supplements– are isolated via the mass

production of a specific bacterial species, Corynebacteria glutamicum. Another example of a

primary metabolite commonly used in industrial microbiology includes citric acid. Citric

acid, produced by Aspergillus niger, is one of the most widely used ingredients in food

production. It is commonly used in pharmaceutical and cosmetic industries as well (Desire et

el., 2014).

2.2.2 Secondary Metabolites

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Secondary metabolites are typically organic compounds produced through the modification

of primary metabolite synthases. Secondary metabolites do not play a role in growth,

development, and reproduction like primary metabolites do, and are typically formed during

the end or near the stationary phase of growth. Many of the identified secondary metabolites

have a role in ecological function, including defense mechanism(s), by serving as antibiotics

and by producing pigments. Examples of secondary metabolites with importance in industrial

microbiology include atropine and antibiotics such as erythromycin and bacitracin. Atropine,

derived from various plants, is a secondary metabolite with important use in the clinic.

Atropine is a competitive antagonist for acetycholine receptors, specifically those of the

muscarinic type, which can be used in the treatment of bradycardia. Antibiotics such as

erythromcyin and bacitracin are also considered to be secondary metabolites. Erythromycin,

derived from Saccharopolyspora erythraea, is a commonly used antibiotic with a wide

antimicrobial spectrum. It is mass produced and commonly administered orally. Lastly,

another example of an antibiotic which is classified as a secondary metabolite is bacitracin.

Bacitracin, derived from organisms classified under Bacillus subtilis, is an antibiotic

commonly used a topical drug. Bacitracin is synthesized in nature as a no ribosomal peptide

synthetase that can synthesize peptides; however, it is used in the clinic as an antibiotic

(Yadav et el., 2014).

2.3 Characteristics of Fungi

Following are the important characteristics of fungi:

i. Fungi are eukaryotic, non-vascular, non-motile and heterotrophic organisms.

ii. They may be unicellular or filamentous.

iii. They reproduce by means of spores.

iv. Fungi exhibit the phenomenon of alternation of generation.

v. Fungi lack chlorophyll and hence cannot perform photosynthesis.

vi. Fungi store their food in the form of starch.

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vii. Biosynthesis of chitin occurs in fungi.

viii. The nuclei of the fungi are very small.

ix. The fungi have no embryonic stage. They develop from the spores.

x. The mode of reproduction is sexual or asexual.

xi. Some fungi are parasitic and can infect the host.

xii. Fungi produce a chemical called pheromone which leads to sexual reproduction in

fungi.

xiii. Examples include mushrooms, moulds and yeast.

2.4 Uses of Fungi

Fungi are one of the most important groups of organisms on the planet as they play a vital

role in the biosphere and have great economic importance on account of both their benefits

and harmful effects.

Following are some of the important uses of fungi:

i. Recycling: They play a major role in recycling the dead and decayed matter.

ii. Food – The mushrooms species which are cultured are edible and are used as food by

humans.

iii. Medicines: There are many fungi that are used to produce antibiotics and to control

diseases in humans and animals. Penicillin antibiotic is derived from a common

fungus called Penicillium.

iv. Biocontrol Agents: Fungi are involved in exploiting insects, other small worms and

help in controlling pests. Spores of fungi are used as a spray on crops.

v. Food spoilage: Fungi play a major role in recycling organic material and are also

responsible for major spoilage and economic losses of stored food.

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18
 Chapter Three

Materials and Methodology

3.1 Materials/Apparatus

3.1.1 Materials

i. Tween 80

ii. Lacto phenol blue

iii. Methanol

3.1.2 Apparatus

i. Sterilized Bags

ii. Potato Dextrose Agar (PDA)

iii. Sterile Spreader

iv. Microscope

v. Conical Flask

vi. Polyethene Bag

vii. Liquid Chromatography

viii. Mass Spectrometer

ix. Perforated Foil Paper

3.2 Collection of Soil Sample

The soil sample were collected from two different locations at Bayero University Kano old

Campus at random within Faculty of Pharmacy. The soil was taken at 5cm depth and inserted

into small sterilized bags for laboratory analysis (Powlson, 2005).

3.3 Identification of Isolated Fungi

A drop of lacto phenol blue was placed in a clean microscope slide, small portion of growth

midway between the colony center and the edge was removed and placed on the slide using

inoculating needle, and cover slip was placed at the edges of the lacto phenol blue, and then

19
mounted on the microscope for examination. Fungi were identified using standard Pictorial

atlas of soil and seed fungi by Watanabe (2001).

3.4 Isolation of Soil Fungi

Two drops of tween 80 was placed in 500ml of distilled water, one gram of soil sample was

then placed in 100ml of the tween 80 solution. The mixture was shaken vigorously, until

homozygous mixture was obtained. Two drops of the homozygous mixture was then placed

in the prepared potato dextrose agar (PDA) and spread using a

sterile spreader. The inoculated plate was sealed and incubated in a room temperature in an

inverted position for about one week, growth of fungi was observed (Powlson, 2005).

3.5 Extraction of Metabolites

The most dominant colony of fungi was removed and transferred into 250ml conical flask.

About 50 ml of methanol was poured, ensuring that all the fungal mycelium is covered. The

conical flask was covered with polyethene which make it air tight, and stored for 5 days. The

methanol was filtered in a fresh conical flask. The methanol extract container was covered

with perforated foil paper. Large pores were made on the foil paper for the methanol to

evaporate and leave the residue for identification (Richard, 2007).

3.6 Identification of Metabolites

The fungal metabolites were identified using LCMS which involves extracting the

metabolites and separating it using liquid chromatography and identified based on their mass

charge ratio detected by the mass spectrometer.

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 Chapter Four

Result and Discussion

4.1 Results

4.1.1 Fungi Isolated From Soil

The microscopic identification of two cultures we’re identified as Pythium Nayoroensa, and

Pythium Irregulare.

Plate 1: Pythium Nayoroensa

Plate 2: Pythium Irregulare

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Table 1: Metabolites Identification in Pythium

Compound Retention Analyzer/Ionization Compound M+H Tentative

Peak Time mode Mass (da)
(Min)
1 QqQ/ESI (+) 169 170.17 Norepinephrine
(Castechols)
2 QqQ/ESI (+) 281 282.31 Oleamide
(Fatty Amides)
3 QqQ/ESI (+) 278 279.3 Gamma-
Linoleic Acid
4 QqQ/ESI (+) 178 179.38 Methylleugenol
5 QqQ/ESI (+) 506 507.29 R1-Barrigenol
(Triterpenoids)
The table above shows that, in Pythium Nayoroensa there were 5 numbers of compounds

sorting base on maximum intensity showed compounds emitting at 10.240 retention time has

molecular weight of 507.29 which corresponds to R1-Barrigenol (Triterpenoids)

Fig 3.1: LCMS Chromatogram in Pythium Nayoroensa

Table 2: Metabolites Identification in Pythium Irregulare.

22
 Compound Retention Analyzer/Ionization Compound M+H Tentative
Peak Time mode Mass (da)
(Min)
1 QqQ/ESI (+) 292 293.29
(+/-)
Altenuene (2-
Benzopyrans)
2 QqQ/ESI (+) 278 279.3 Gamma-
Linoleic Acid
The table above shows that in Pythium Irregulare were 2 numbers of compounds sorting

base on maximum intensity showed compound emitting at 10.283 retention time has a

molecular weight of 279.3 corresponds to Gamma-Linoleic Acid.

Fig 3.2: LCMS Chromatogram in Pythium Irregulare

4.2 Discussion of Results

The genus Pythium is one of the most important groups of soilborne plant pathogens,

present in almost every agricultural soil and attacking the roots of thousands of hosts,

reducing crop yield and quality. Most species are generalists, necrotrophic pathogens that

infect young juvenile tissue. In fact, Cook and Veseth (29) have called Pythium the

“common cold” of wheat, because of its chronic nature and ubiquitous distribution.

23
Many diseases caused by Pythium spp. are favored by wet, cool conditions, although some

species are more commonly encountered under warmer conditions.

In addition to examining populations of pathogenic species of Pythium, real-time PCR has

also been used to quantify non-pathogenic isolates of Pythium being used for biological

control.

Plant-associated fungi are incredibly diverse, comprising over a million species through

world wide. This diverse fungal community is highly important for plant health. Many fungi

are effective biocontrol agents that can kill or suppress fungal pathogens, with pathogen

biocontrol found for both individual microorganisms and plant-associated fungal consortia.

Meanwhile, increased plant community diversity aboveground corresponds to an increase in

below-ground fungal community diversity, which contributes in turn to improved soil health

and pathogen suppression.

24
 Chapter Five

Summary, Conclusion, Recommendation and References

5.1 Summary

Soil fungi play a crucial role in various ecological processes, including nutrient cycling and

decomposition. They also produce a diverse range of metabolites that have potential

applications in medicine, agriculture, and other industries. This project aims to assess the

metabolites produced by soil fungi and explore their diversity, functions, and potential

applications.

5.1 Conclusion

In Conclusion, the soil analyzed contains different types of fungal species. The two common

soil fungi obtained were Pythium Nayoroensa, and Pythium Irregulare. Extraction of the

fungal metabolite from the crude extract using LC-MS revealed the presence of many

compounds among which Pythium Nayoroensa shows the molecular weight which

corresponds to R1-Barrigenol (Triterpenoids) and Pythium Irregulare which has a molecular

weight that corresponds to Gamma-Linoleic Acid. To conclude, the assessment of

metabolites in soil fungi is crucial for understanding their ecological roles, interactions with

other organisms, and potential applications. Advanced analytical techniques and further

research are necessary to explore the diversity and functions of fungal metabolites in various

soil ecosystems. By gaining a deeper understanding of these metabolites, we can harness their

potential benefits and mitigate their harmful effects, contributing to sustainable agriculture,

environmental health, and human well-being.

5.2 Recommendations
25
1. Advanced analytical techniques and further research are necessary to explore the

diversity and functions of fungal metabolites in various soil ecosystems.

2. Further research shall be carried out to explore the vast metabolites locked up in

fungal species and to utilize them in various field of human lives.

26
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