Morpho - anatomical and molecular

analysis of different basidiomycetes

Researcher:
Salma Javaid
Roll No: K3F18ASBZOO78
BS(ADS Program 2 years)

Supervisor:
Prof Syed Baqir Latif

10
 “In The Name Of Allah, The Most Gracious, The Most
Merciful. The Most Lenient.”

“Praise is to Allah, lord of the Worlds, the

Beneficent, and the Merciful Owner of the Day of Judgment.
Thee (alone) we worship; Thee (alone) we ask for help. Show 11
us the straight path, the path of those whom thou has favored;
not (the path) of those who earn thine anger nor of those who
go astray.”
 (Surah AL-Fateha)

Morpho - anatomical and molecular analysis

of different basidiomycetes

By
SALMA Javaid

Under the Supervision of

Prof. Syed Baqir Latif
B.s(Botany)

A thesis submitted to University of The Central Punjab in the partial fulfillment of

requirements for the degree of B.s Botany.

UNIVERSITY OF CENTRAL PUNJAB, 12

LAHORE-PAKISTAN
SESSION-2020
 DEDICATED

To

“MY PARENTS”

&

“TEACHERS WHO HAVE HELPED US IN

THE COMPLETION OF THIS
HARDWORKING JOB”

13

CERTIFICATE OF APPROVAL
 It is certified that this thesis is based on the results of work done by
_(Salma Javaid . I certify that all the data and results reported in the thesis are correct
and authentic. __Salma Javaid __ .fulfilled all the condition of University of Central
Punjab for the submission of this thesis. It is hereby approved for submission to the
University of The Central Punjab, Lahore for the partial fulfillment of the requirement for
the degree of BS in Botany.

Supervisor Name:———————— Signature: —————————

Date:————————

14

Declaration
This is to certify that this research work has not been submitted for obtaining similar
degree from any University College.

Prof.Syed Baqir Latif

2020

15

COURSE COMPLETION CERTIFICATE

 It is certified that Course Requirements of Salma Javaid ROLL NO:
K3F18ASBZOO78 for the B.S (Botany) Program has been complete.

Prof. Syed Baqir Latif

16

Research Completion Certificate

 It is certificated that the research work contained in this dissertation
entitled “Effectiveness of teacher diary in teaching learning process” has been carried out
and completed SALMA JAVAID Under the Supervision during her B.S Botany
Programmed.

Supervisor

Prof. Syed Baqir Latif

17
 Acknowledgements
I would like to express my gratitude to my supervisor Prof.Syed Baqir Latif Without
his valuable advice and encouragement the completion of this thesis would not have been
possible. I am deeply grateful to my mother whose prayers help me in my study. I am
thankful to my family and siblings. I am also thankful to my friends who guide me in
each step.

Table content :

Anstract

 Basidiomycetes
 Order
 Introduction
 Basidiomycetes based on ITS-rDNA
 Methods and materials
 Keywords
 Morphological variations of basidiomycetes18
 Ecology of marine basidiomycetes
  Basidiomycetes general characters
 Life cycle
 Types of basidiomycetes
 Importance of Basidiomycetes
 Uses of basidiomycetes

Abstract :
Basidiomycetes are a diverse assemblage of
fungi represented by over 20,000 species
worldwide. They include the familiar, agaris,
boletes, club and coral fungi, chanterelles,
jelly fungi, polypores, puffballs and tooth
19

fungi, and the not so familiar

basidiomycetous yeasts. They obtain their
nutrition as saprotrophs, pathogens and
mutualists, and play an important role in
ecosystem functions. Basidiomycetes are
abundant and diverse in Thailand although
they are currently poorly known. We
estimate that the approximately 300 species
reported in the literature from Thailand
represent less than 20% of the actual
diversity.

Basidiomycetes :
The class Basidiomycetes includes those
members that produce their basidia and
basidiospores on or in a basidiocarp. The
morphology of the basidium is variable. Until
recently the morphology of the basidium was
believed to be a key to determining relationship
20

in the Basidiomycota. Basidial morphology was

once the basis for classifying the fungi to class
or subclass. However, rDNA sequencing analyst
(Swann and Taylor, 1993), septal pore
morphology and cell wall biochemistry
(McLaughlin et al, 1995) have determined that
far too much emphasis was placed on this
characteristic and all members of the
Basidiomycota that produce basidiocarps are
now included in a single class, the
Basidiomycetes, and the morphology of the
basidiocarp and basidium are characteristics that
are now used to classifying fungi into the
various orders of this class.
Order :
Agaricales

This is the order of Basidiomycetes with which

most of us are familiar. This is the order that is
commonly referred to as mushrooms.
Basidiocarps of this order typically are "fleshy"
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and have a stipe (=stalk), pileus (=cap), and
lamellae (=gills) where the basidia and
basidiospores are borne (Fig. 1-4). The
Basidiospores in this order of fungi are forcibly
ejected from the basidium, into the area between
the lamellar edges, which then allows the spores
to fall free from the mushroom and be dispersed
by wind.
Molecular analysis of different
basidiomycetes based on ITS - rDNA:
Lepiota acutesquamosa and L. cristata
(Basidiomycota, Agaricales) collected from
Himalayan moist temperate forests of Pakistan
were characterized using internal transcribed
spacers (ITS) of rNDA, a fungal molecular
marker. The ITS-rDNA of both species was
analyzed using polymerase chain reaction
(PCR) and DNA sequencing. The target region
when amplified using universal fungal primers
(ITS1F and ITS4) generated 650-650bp
fragments. Consensus sequences of both species
were submitted for initial blast analysis which
revealed and confirmed the identification of 22
both species by comparing the sequences of
these respective species already present in the
GenBank. Sequence of Pakistani collection of L.
acutesquamosa matched 99% with sequences of
same species (FJ998400) and Pakistani L.
cristata matched 97% with its sequences
(EU081956, U85327, AJ237628). Further, in
phylogenetic analysis both species distinctly
clustered with their respective groups.
Morphological characters like shape,size and
color of basidiomata, basidiospore size, basidial
lengths, shape and size of cheilocystidia of both
collections were measured and compared. Both
these species have been described first time
from Pakistan on morph-anatomical and
molecular basis.
Introduction :
Basidiomycota belong to the crown group of fungi.
The diversity of cellular constructions in hyphal
systems and basidiocarps is the expression of a long
evolutionary history. As heterotrophic Eukaryotes
these fungi developed substrate dependencies of 23
enormous ecological importance, especially in
forest ecosystems.
Methods and materials :
Samples tested were prepared by hot water
extraction from various wild Basidiomycetes, which
were gathered in various parts of Japan. Since the
samples were hygroscopic, they were lyophilized in
order to keep them sufficiently dry.
Keywords :
Diversity, fungi, taxonomy.
Morphological variations in basidiomycetes :
Oidiophores with uninucleate oidia are produced on
the aerial mycelium of Coprinus cinereus and
occasionally also on the submerged mycelium.
Oidiophore development starts with the formation
of a stem cell and proceeds with stem cell
elongation, followed by the formation of oidial
hyphae at the tip of the stem cell and subsequent
release of matured oidia. However, analysis of more
than 20 different C. cinereus strains revealed that
oidiophore formation is a flexible process. Based on
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morphological variations, we defined four main
types of oidiophores. Types 1 and 2 oidiophores
produced oidia at the tip(s) of simple (type 1) or
branched stems (type 2). Types 3 and 4 oidiophores
were characterized by the absence of stem cell
elongation (type 3) or stem cell formation (type 4).
All strains examined produced types 1 and 2
oidiophores and some produced also types 3 and 4
oidiophores but the frequency of the different
oidiophore types varied strongly from strain to
strain.
Ecology of marine basidiomycetes :
Marine and freshwater basidiomycetes are few in
number compared to their terrestrial counterparts
and colonize a wide range of substrata: sea-grasses,
feathers, wood associated with sand, free floating in
the sea, but most occur on mangrove wood or
timbers submerged in the sea (boats, piling, sea
defences), and leaves and twigs in streams and
rivers. They are an ecological group and
taxonomically diverse (Agariomycotina,
Uredinomycotina and Ustilaginomycotina). Most
are able to utilize simple carbohydrates, while
filamentous species can decompose cellulose, 25

hemicellulose and lignin. Aquatic basidiomycetes

are well adapted to their habitats, with reduced
basidiomata. Marine species are known only as
teleomorphs with basidiospores generally released
passively. Freshwater basidiomycetes are primarily
known by their anamorphs on decaying leaves, with
conidia that are much branched, while their
teleomorphs occur on land and on woody substrata.

Basidiomycetes general characters :

The Basidiomycetes comprise the highest group of
fungi which are often noticed conspicuously
occurring in fields and forest areas. Most of them
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possess highly developed fructifications, known as
basidiocarps which may be fleshy or woody.
The fungi commonly known as mushrooms,
toadstools, puff-balls, and bracket fungi are the
Basidiomycetes. To this group also belong the rusts
and smuts, responsible for diseases of plants of
economic importance. There are both parasitic and
saprophytic fungi in this group.
Mating systems in basidiomycetes :
About 10 per cent, of Basidiomycetes are
homothallic.
Three types of homothallic behaviour may be
distinguished:

(i) Primary homothallism:

In Coprinus sterquilinus, a single basidiospore
germinates to form a mycelium which soon
becomes organized into binucleate segments
bearing clamp connections at the septa. There is no
genetical distinction between the two nuclei in each
27

cell, and mycelium is capable of forming fruit

bodies.
(ii) Secondary homothallism:
In Coprinus ephemerus f. bisporus, the basidia bear
two spores, but the spores are heterokaryotic. After
meiosis two nuclei enter each spore and a mitotic
division may follow. On germination, a single spore
germinates to form a dikaryotic mycelium capable
of producing fruit bodies.

(iii) Unclassified homothallism:

The four-spored wild mushroom Agaricus
campestris and the two-spored cultivated
mushroom Agaricus bisporus are homothallic in the
sense that a mycelium derived from a single spore is
capable of producing fruit bodies. In both species
there is nuclear fusion in the basidium, followed by
two nuclear divisions, presumably meiotic.
Life cycle of basidiomycetes :
The general life-cycle pattern of the Basidiomycetes
has resemblance with that of the Ascomycetes. The 28
basidiospore on germination by germ tube gives
rise to the haplophasic somatic body represented
by primary mycelium. The primary mycelium often
produces oidia.
The oidia may behave as spores and give rise to
primary mycelia, or by spermatization with
compatible primary mycelia take part in the
formation of dikaryotic secondary mycelia. The
secondary mycelium may also be formed by
somatogamy between two compatible primary
mycelia. The dikaryotic apical cell of the hypha of
the secondary mycelium develops into basidium.

It may so happen that resting spores may be formed

from the dikaryotic hyphal cells. Each resting spore
on germination gives rise to a basidium. Karyogamy
takes place in the basidium producing diplophasic
condition which is followed immediately by meiosis
and ultimately haploid basidiospores are produced.
The basidiospores germinate to produce
haplophasic somatic body.
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Thus the life cycle is completed. The life cycle

comprises
of the three phases the haplophase, dikaryophase
and diplophase; of which the dikaryophase is very
much prolonged and is nutritionally independent of
the haplophase.

Types of basidiomycetes : 30

More specifically, Basidiomycota includes these

groups: mushrooms, puffballs, stinkhorns, bracket
fungi, other polypores, jelly fungi, boletes,
chanterelles, earth stars, smuts, bunts, rusts, mirror
yeasts, and the human pathogenic yeast
Cryptococcus.
Mushroom :

Puffball mushroom :

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Importance of Basidiomycetes :
Economically, mushrooms are valued for food and
for bioactive compounds exploited in traditional
medicine. Wood decay Basidiomycetes can destroy
construction timber, but are valuable in
lignocellulose conversion, for composting and as
potential producers of biofuels. Rusts and smuts
cause major crop losses. 32

Used for :
The bastidiomycota help the plants obtain
nutrients from the soil, and in return recieve
sugars produced through photosynthesis.
Bastidiomycota also form symbiotic relationships
with insects. While mushrooms are used for food,
there are also many other uses humans have for
Basidiomycota
Economic importance of Basidiomycetes :
Many of the Basidiomycetes are of great economic
importance because of their beneficial as well as
harmful nature. Some of them are the causative
agents of most destructive diseases of our cereal
crops.

To this category belong the smut diseases of com,

wheat, oats, and barley as well as the wheat rusts.
They destroy several million rupees worth of crops
every year.
Many of the Basidiomycetes are of great economic
33
importance because of their beneficial as well as
harmful nature. Some of them are the causative
agents of most destructive diseases of our cereal
crops.

To this category belong the smut diseases of com,

wheat, oats, and barley as well as the wheat rusts.
They destroy several million rupees worth of crops
every year.
Reproduction in basidiomycetes :
Image of Basidiomycota reproduction
Image of Basidiomycota reproduction
The phylum Basidiomycota is a group of fungi
characterized by the formation of specialized club-
shaped cells, called basidia, during reproduction.
The basidia normally produce four haploid spores,
called basidiospores. Some Basidiomycota
reproduce asexually, and some reproduce sexually.
Sexual reproduction in basidiomycetes ;
Sexual reproduction in Basidiomycota takes place in
34
the fruiting body, in specialized structures called
basidia. The basidia is itself formed by plasmogamy
between mycelia from two different spores. ...
These projections are then separated by cell walls
to become spores.
Asexual reproduction in basidiomycetes :
Asexual Reproduction
Basidiomycota reproduce asexually by either
budding or asexual spore formation. Budding
occurs when an outgrowth of the parent cell is
separated into a new cell. ... The septae of
terminal cells become fully defined, dividing a
random number of nuclei into individual cells.
Unlike most fungi, basidiomycota reproduce
sexually as opposed to asexually. Two different
mating strains are required for the fusion of genetic
material in the basidium which is followed by
meiosis producing haploid basidiospores.
Basidiomycota are filamentous fungi that are made
of hyphae (except for yeasts). They reproduce
sexually by the formation of specialized club-shaped
cells called basidia. The basidia normally produce
35
four haploid spores, called basidiospores. Members
of the phylum Basidiomycota.
Club basidiomycetes reproduction :
Species in this phylum reproduce sexually by
forming spores on top of club-shaped structures
called basidia. The club fungi are believed to be
closely related to the sac fungi. ... The club fungi
reproduce asexually by producing asexual spores or
by fragmentation of mycelium.
Basidiomycetes structure :
Basidiomycota are filamentous fungi composed of
hyphae (except for basidiomycota-yeast; refer yeast
for more information) and reproduce sexually via
the formation of specialized club-shaped end cells
called basidia that normally bear external
meiospores (usually four).

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U85327

37

Basidiomycetes taxonomy :
Basidiomycota is the monophyletic phylum that
contains Chlorophyllum molybdites along with
many others. Basidiomycota is broken down into
many different classes, orders, families, genus', and
species.

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