SAN CARLOS UNIVERSITY OF GUATEMALA

UNIVERSITY CENTER OF THE WEST

DIVISION OF SCIENCE AND TECHNOLOGY
GENERAL MICROBIOLOGY LABORATORY

MICROSCOPIC PREPARATIONS OF FUNGI

Fungi comprise the largest group of phytopathogenic microorganisms,

also being the causes of the largest agricultural economic losses, due to the
a large number of diseases they cause.

It is considered that all plants are susceptible to the attack of at least one fungus.
and many are affected by a large number of these organisms.

The habitat of fungi is very broad, as they can be found in the soil, in the water and
inside or about plants and animals. They can develop in very climatic conditions
varied, finding them in temperature ranges from 0ºC to 50º.

Fungi belong to the kingdom Fungi, they are microscopic and macroscopic, uni and
multicellular, lacking chlorophyll. They are made up of round or oval cells.
solitary or in plasmodia, or more frequently by elongated and branched structures
hyphae, whose set is called mycelium. Each hypha is made up of a
cell wall that protects the cell membrane and the protoplasmic content, where
their true nuclei are dispersed, as well as the mitochondria, ribosomes,
endoplasmic reticulum and storage substance granules. If the hyphae are separated
in cells, it is said that the mycelium is septate, and if the protoplasm is continuous in the hyphae
The mycelium is coenocytic. In some fungi, the mycelium can form pseudotissues.
Prosenchyma and pseudoparenchyma.

Among the structures of asexual reproduction are oidiums, chlamydospores,

sporangia y conidia, the latter solitary o grouped in cinemas,
esporodoquios, acervulos or picnidios. Some structures of sexual reproduction are the
asci, free or in apothecia, perithecia and cleistothecia, and basidia, that arise
directly from a resistance spore or in basidiocarps.

CHARACTERISTICS TO IDENTIFY FUNGI

Macroscopic aspect of the colony
Type of hypha
Placement of the or thesporangia
Presence ofesterigmatasporangiophoreoconidiophoreand the order they present
Shape, size, and distribution of the spores
Presence or absence [Link] only appear in non-septate hyphal fungi.
example:Rihizopus, Rhizomucor,Absidia
Practice biochemical identification tests.
 CLASSIFICATION OF FUNGI

Zygomycotina: In the zygomycotina, two typical hyphae of these, which are multinucleated.
They can contact and if they are complementary, they will partition the portions at the end that remain.
constituted as gametocysts. The gametocysts fuse (plasmogamy) but do not
Immediately after karyogamy, the nuclei are paired. After a while,
At times the nuclei fuse (karyogamy) forming a polyzygot or zygospore that
is surrounded by a strong wall, entering a state of resistance. The
ascomycotina and basidiomycotina have clear sexual reproduction, which makes them
difference of the deuteromycotina, whose sexual reproduction is unknown. Among the
Zygomycetes, perhaps the best known are some representatives of the order Mucorales.
likeMucoroRhizopus, but in reality the group has members that represent
very disparate evolutionary lines. They are characterized by amyceliumacceptedcenocytic
with septa at the base of reproductive structures or secondary septa. The name
of the group comes from the presence in part of its cycle of azigosporecharacteristic.
But in the entire group, asexual reproduction is also very important, even
Some classification systems of group parts are based on this. Within this
we have a very important order agronomically, that of the Glomales,
they will be addressed in the agendamycorrhizas.

Ascomycotina: The ascogonium and the antheridium are the gametocysts, male and female.
respectively, of the ascomycetes. These gametocysts differentiate at the ends
of some hyphae. In fungi, we speak of homothallic species (which are equivalent to
monoecious species) and heterothallic species (which correspond to dioecious species). In
ascomicetes, can there be a case where there are no differences in
gametocysts. The male role is usually developed by a cell called spermatozoid,
that will contact a terminal cell of a hypha that will act as a female gamete,
At first, the mycelium is monokaryotic. First, fusion occurs.
from the gametocysts (plasmogamy), but it is not immediately followed by the fusion of
nuclei, since these nuclei pair up. Then the onset of the mycelium occurs.
dikarotic that presents cells with two complementary nuclei that do not fuse.
There are fungi where the monokaryotic phase is dominant, but in others, the dikaryotic phase is.
dominates, even the monokaryotic phase is very fleeting, as two spores contact and
the dikaryotic mycelium quickly forms. The two complementary nuclei synchronize.
its division, in such a way that it allows for the orderly organization of the mycelium. The ends
from the cells of the dikaryotic hyphae form a kind of hook, the uncinulus. Each
the nucleus of the uncinate cell synchronizes its divisions, they divide in such a way that in
At each end, there are two complementary cores separated by a partition, thus
Well, we have two complementary cores at the top and another two at the bottom. Between
both the uncino has just contacted the hifa, and the wall that separates the two nuclei
the inferior disappears, that is, the uncinate disappears. This process repeats until
the main cell decides to fuse the two nuclei (karyogamy), resulting in the cell
diploid. In fungi, the concept of haploid and diploid is not usually used, but rather it
talks about monokaryotic mycelium and dikaryotic mycelium. In this example, plasmogamy and the
cariogamia are very separated in time and space. Next will take place
meiosis and the successive mitoses until having 8 haploid cells that develop a
cover and give rise to spores. With the process of division, the cell enlarges until
to constitute a sexual-origin spore cyst known as an ascus. The spores are
they are called ascospores and are endogenous. When a spore germinates in species
homotallic or spore-forming in heterotallic species, there is the development or formation of the
monokaryotic mycelium that progresses to the formation of gametangia, which
They are contacted through the trichogyn. The covering of the ascospores is called the tunica.
There can exist proto-tunics which are very poorly organized allowing to let out
All around the ascospores. The tunics can be unitunicate when the fall of
the ascospores are produced at the end, sometimes they have a lid (operculated) by the
that the ascospore falls, or simply the end breaks (inoperculate). Another type
The tunics are the bitunics when a double tunic is presented. The ascas
They generally contain 8 ascospores, but there are some asci that contain four.

Basidiomycotina: The fungi belonging to this subdivision have the body

fruiting on the surface, these fruiting bodies are the carriers of the spores.
To study the mode of reproduction, we start from two spores, so we have a
heterotalic species, which organizes a monocarlic mycelium (septate cells that
they contain a single nucleus). Unlike in ascomycotina, it does not end in any
special structure (there are no special gametocysts). The cells act as
gametocysts join and plasmogamy occurs, but fertilization does not occur immediately
Dikaryogamy. The dikaryotic mycelium organizes from this moment through clamps.
The fibula is very similar to the hook, but appears laterally. One of the nuclei is
it is located in the fibula and divides at the same time as the other nucleus. The septa appear,
one is located in the loop formed by the fibula and ultimately contacts the nucleus
complementary, the dicarious mycelium is progressing. The basidiomycetes,
Generally, they can be differentiated because they have handles, although this is not the case for all.

the cases. The dikaryotic mycelium divides very profusely until it forms bodies
fruitful. Sometimes the dikaryotic mycelium can be accompanied by some portion of
monokaryotic mycelium that provides protection and support to the dikaryotic ones. The mycelium
monocaryotic ends in an infertile structure, the paraphysis.
In basidiomycetes, the fruiting body is called a basidiocarp, whereas in
In ascomycetes, the fruiting body is called an ascocarp. The important part of the
The basidiocarp is the cap, where the fertile part or hymenium is located, which is constituted by
by the set of basidia (sporocysts of sexual origin in basidiomycetes). The hymenium
coat the laminas. The cell at the end of a dikaryotic mycelium is the one that decides the
fusion of the nuclei (karyogamy) to form the zygote, which is the only diploid cell.
This cell grows larger and becomes the basidium, while inside, that cell
diploid undergoes meiosis that is not followed by mitosis. Thus we have four
nuclei that migrate to small protrusions where they are located and at the same time they
these projections constrict, so that the four basidiospores are formed.
The general case is that the basidium is unicellular, without septa (holobasidiomycetes). There is another
group that includes (fragmobasidiomycetes). On the surface of the gills of the fungi
there is a great concentration of basidia. Many times these basidia do not reach a good
term and are called basidioles or cystidia, which have the mission of attracting the
insects o that the basidia do not become compacted. They are plectenchymatous.o
pseudoparenchymatous, the arrangement of the hyphae forms characteristic traces, these
traces finish superficially in the hymenium where different types of cells converge.
Some ascomycetes also have fruiting bodies, but the typical ones are from the
basidiomycetes.

Deuteromycotina: In this subdivision of fungi, phenomena can be found.

parasexuality in some representatives. These processes successfully replace and
equivalent to the sexuality processes of ascomycetes and basidiomycetes. They take place
processes of recombination and the emergence of combinations without resorting to processes of
sexuality. It has been found in a few species. Some yeasts have a
independent DNA strand, the plasmids. Some very simple ascomycetes have
very small DNA structures that contain genetic information independent of
contained in the nucleus or in the mitochondria. This fact may be related to the
resistance that some yeasts exhibit against antibiotics. Plasmids have
few nucleotides are encoded, but they are enough to help them in their defense.
 ACTIVITIES.

Preparation of temporary mushroom preparations.

2. Observation of permanent preparations.

MATERIALS AND METHODS.

Slide and cover slip
Lactofenol
Distilled water
Petri dish
Dissection needles
Mushroom preparations
Sick plant tissues

Temporary preparations of fungi

1. With a sterile loop (or a dissecting needle), take a small sample of fungi.
and place it on a slide with a drop of distilled water.
2. Place the coverslip and observe under the microscope.
3. Identify the fungus and its parts.
4. Note the observations and make comparisons of the different fungi observed.

Make temporary preparations with clear adhesive tape and observe them.
microscope.

PERMANENT PREPARATIONS OF FUNGI

1. Place the permanent preparations under the microscope and identify the parts of the
fungus and the name of the observed fungus.

QUESTIONNAIRE:

What does the word fungi mean?

2. What cellular organisms are in the kingdom fungi?
3. Are fungi prokaryotes or eukaryotes:
4. How do fungi reproduce?
5. What emerges after the spores germinate?
Fungus is used in the production of antibiotics.
7. How is the kingdom fungi classified?