Principles of Biology 2/11/15

Fungi
Chapter 31 (Campbell, 10th ed.)

The origin of Fungi

Kingdom closest to animal kingdom

Choanoflagellates
Animals
Unikonta
Fungi
Amoebozoans

Fungi and animals diverged about 1.5 billion years ago

Robert Whittaker (1920-1980)

Fungi represent a separate kingdom (1959)

The Quarterly Review of Biology, Vol. 34, No. 3 (Sep., 1959), pp. 210-226

Types of fungi
>100,000 species described
Yeasts

Unicellular microscopic fungi

Not a phylogenetic group

Molds

Filamentous microscopic fungi

Not a phylogenetic group

Mushrooms

Macroscopic fungi
Not a phylogenetic group

Impact of Fungi on the world we live in

Breakdown of organic material, recycle nutrients

(major ecological role)
Produce antibiotics (penicillin)
Can cause diseases

Fungal pathogens
~300 species of fungal pathogens

Cause diseases:
Toxin production (mycotoxins)
Infections (mycoses)
Allergic responses

Tintin: The Shooting Star

Story of Babar

Mycotoxins
Aflatoxin B1

Ergotamine

Carcinogenic, liver disease

Ergots (Claviceps purpurea) on rye
Aspergillus flavus on corn

Source: NCSU

Ergotism
Symptoms: hallucinations, convulsions and gangrene (by vasoconstriction)
Saint Anthonys fire

Temptation of St Anthony
by Matthias Grunewald (~1515)

Salem witchcraft trials?

Science, vol. 192, 1976

Science, vol. 194, 1976

Le pain maudit: the cursed bread

1951 Pont Saint-Esprit, France (200 cases, 4 deaths)

Lysergic acid diethylamide (LSD)

LSD

Albert Hofmann
(1906-2008)

Mycoses
Superficial

surface layers (hair, skin, or nails)

Subcutaneous

deeper layers of skin

Systemic

fungal growth
in internal organs of the body

f
Romani (2004)

Fungi are heterotrophs that feed by absorption

Fungi do not ingest food and are not phototrophic

Large molecules are broken down into smaller compounds that can be absorbed

Fungi grow next to their main source of nutrients

(most fungi are not motile, except some flagellated fungi)

Exoenzymes

hydrolytic enzymes secreted in the environment

(food is digested outside of the body)

Body structure
Hyphae (=tubular filaments)

Hyphae

Hyphae

Spore-producing
structures

60 m
Mycelium

Mycelium (=network of hyphae)

surrounds and infiltrates the material on which the fungus feeds

Structure of hyphae
Cell wall contains chitin
=polymer of NAG (N-acetyl glucosamine)
Cell wall

Cell wall

Nuclei
Pore
Septum

(a) Septate hypha

Higher fungi

Nuclei

(b) Coenocytic hypha

Lower fungi

Fungal reproduction
Asexual
Simple cell division
Growth and spread of hyphal filaments
Asexual production of spores

Sexual
Sexual production of spores

Spores can be produced either sexually or asexually (but are always haploid)

Lifestyles
Fungi get nutrients from all sorts of sources

Decomposers (saprobes)

Parasites

Mutualistic symbionts

dead organisms or waste from living organims

Spores

Fungi use spores to propagate themselves

Fungal life cycle

Spores

Spore
germination

Mature
mushroom

Spores are resistant to drying, heating, freezing, chemicals

2012 Pearson Education, Inc.

Sexual reproduction of Fungi

Different mating types
(may appear identical morphologically but chemically different)
Pheromones
(=sexual signaling molecules, e.g. a and )

Hyphae or cells (yeast) extend toward the source of pheromone and fuse

Generalized life cycle of fungi (sexual)

Unfused 2n stage

Key
Haploid (n)
Heterokaryotic
(unfused nuclei from
different parents)
Diploid (2n)

Heterokaryotic
stage
PLASMOGAMY

KARYOGAMY
SEXUAL
REPRODUCTION
Mycelium

In fungi, 2 stages

Haploid
(n)

Syngamy

fusion of gametes

1) Plasmogamy

fusion of cytoplasms

2) Karyogamy

fusion of nuclei

Zygote

Diploid
(2n)

Generalized life cycle of fungi (sexual)

Key
Haploid (n)
Heterokaryotic
(unfused nuclei from
different parents)
Diploid (2n)

Heterokaryotic
stage
PLASMOGAMY

KARYOGAMY
SEXUAL
REPRODUCTION

Zygote

Mycelium

MEIOSIS
GERMINATION

Spores

Spore-producing
structures

Generalized life cycle of fungi (asexual)

Key
Haploid (n)
Heterokaryotic
(unfused nuclei from
different parents)
Diploid (2n)

Heterokaryotic
stage
PLASMOGAMY

KARYOGAMY

Spore-producing
structures

Spores

ASEXUAL
REPRODUCTION

SEXUAL
REPRODUCTION

Zygote

Mycelium

MEIOSIS
GERMINATION

GERMINATION

Spores

Spore-producing
structures

Fungi with no known sexual stage are defined as deuteromycetes or imperfect fungi

Phylogeny of fungi
Chytrids

Zygomycetes

Glomeromycetes

(~1,000 species)

(~1,000 species)

(~150 species)

Ascomycetes
(~65,000 species)

Basidiomycetes

(~30,000 species)

Chytrids
Chytrids

(~1,000 species)
Diverged earliest in fungal evolution
Most chytrids species are motile with flagella
Like all fungal species:

cell walls are made of chitin

form hyphae
Globular fruiting body

Zoospores (flagellated spores)

4 m
Flagellum

Hyphae

Zygomycetes
(~1,000 species)

Zygomycetes

Make zygosporangia

Known primarily for food spoilage

Commonly found in soil and decaying plant material
All are coenocytic
All form zygospores

Rhizopus solonifer
black bread mold

Asexual life cycle of Rhizopus

Genetically identical spores

Sporangium
Dispersal

Germination
Mycelium
Coenocytic hyphae

Sexual life cycle of Rhizopus

PLASMOGAMY

Septation
Mating
type (+)

Mating
type ()

Germination
Zygosporangium
Dispersal

Genetically diverse spores

KARYOGAMY

Diploid
nuclei
Sporangium
MEIOSIS

(heterokaryotic)

highly resistant
to freezing or drying

Genome sequence of Encephalitozoon cuniculi,

a microsporidian
Unicellular eukaryotic parasite that lacks mitochondria

10 m

Microsporidia are zygomycetes

Host cell nucleus

Developing microsporidian

Spore
Keeling (2001)

2.9 Mb; 11 chromosomes; 1,997 genes

Not a primitive eukaryote
but lost most of its morphological characteristic as a result of its parasitic life style
Katinga et al. (2001)

Glomeromycetes
(160 species)

Symbionts with plants

Mycorrhizae= mutually beneficial relationship between fungi and plant roots
Ectomycorrhizae (ectos, out)= hyphae over the surface of the root
Endomycorrhizae (endo, in)= invaginations inside the root cell wall
Glomeromycetes

2.5 m

Plant root

Arbuscular endomycorrhizae
=branched hyphae (tree-like structure)

Specialized hyphae of glomeromycetes

Fungal hypha

Plant
cell
wall

Plant cell
Haustorium

Plant cell
plasma
membrane

Haustoria (specialized hyphae that penetrate the root cell wall)

Having mycorrhizae is beneficial to plant growth

Soybean plants
+ fungicide

no fungicide

no mycorrhizae

mycorrhizae

P deficiency

Healthy plant

Ascomycetes (sac fungi)

Ascomycetes

(~65,000 species)

Aleuria aurantia
(orange peel fungus)

Saccharomyces cerevisiae

Morchella esculenta
(morel)

Neurospora crassa

Tuber melanosporum
(truffle)

Penicillium

Production of sexual spores in sac-like (pod-like) structures termed asci

Sexual stages in fruiting bodies called ascocarps

Ascomycetous yeasts

The budding yeast Saccharomyces cerevisiae

(bakers yeast, brewers yeast)
10 m

Asymmetric division
buds

bud scars
(previous
divisions)

Flourish in habitats where sugars are present

Grows in simple media (C source + salts)
Short generation time ~ 2 hours
Can grow aerobically or anaerobically
Most intensively studied eukaryotic model

Life cycle of S. cerevisiae

S. cerevisiae is a great genetic tool

Although asexual reproduction is favored, S. cerevisiae is able to reproduce sexually
Lethal mutations can be studied in haploids
and maintained in diploids

Foreign DNA can be introduced into yeast cells by electroporation

and integrated by homologous recombination

Yeast artificial chromosomes (YACs)

Eukaryotic proteins can be expressed with correct post-transcriptional modifications

Nobel prize for physiology and medicine, 2001

Leland Hartwell (S. cerevisiae), Tim Hunt (Xenopus oocyte), Paul Nurse (S. pombe)
Discovery of key regulators of the cell cycle
INTERPHASE
S
(DNA synthesis)

C
M yto
ito ki
si ne
s s

is

G1

G2

MI
(M TOT
) P IC
HA
SE

Isolation of temperature sensitive yeast mutants blocked at specific stages of the cycle

Genomes of ascomycetous yeasts

Saccharomyces cerevisiae
budding yeast
first eukaryotic genome sequenced (1996)
12 Mb; 16 chromosomes; 6,275 genes

Schizosaccharomyces pombe

fission yeast, symmetric division, (the other yeast)

diverged from budding yeast ~400 million years ago
genome sequence (2001, 6th eukaryotic genome sequenced)
14 Mb; 3 chromosomes; 4,824 genes
large majority of genes have introns

Candida albicans
Pathogen, skin and mucosal infections (candidiasis)
15 Mb; 7 chromosomes; 6,419 genes
Dimorphic = yeast inside the host, filamentous outside

Ascomycetous molds

Conidia (or conidiospores)

asexual spores formed
at the tip of specialized hyphae
(conidiophores)

Aspergillus
Aspergillus nidulans
Aspergillus fumigatus
Aspergillus oryzae

model organism
human pathogen and allergen
food fermentation (sake, soy sauce)

Aspergillus forms long chains of conidia on globelike conidiophores

Penicillium
Deuteromycetes (imperfect fungi): no sexual cycle

1.5 m
2.5 m

mold

Penicillium forms long chains of conidia on branching conidiophores

Penicillium notatum

Penicillin

Penicillium glaucum

Gorgonzola cheese

Penicillin inhibits transpeptidation

i.e. inhibits formation of cross-links in peptidoglycan
Staphylococcus

Penicillium
Zone of
inhibited
growth

Most-widely used antibiotic to date,

Revolutionized medicine

Two enzymatic activities:

Transglycosylation + transpeptidation
Peptidoglycan
Transglycosylase
activity

Cytoplasmic
membrane
Out

Growing point
of cell wall

Autolysin
activity

In
Pentapeptide

Transpeptidation

Bactoprenol

Use of antibiotics

-lactams (cephalosporins and penicillin)

represent more than half

Discovery of penicillin

Alexander Fleming
(1881-1955)
Serendipitous discovery (1928)

Howard Florey
(1898-1968)

Ernest Boris Chain

(1906-1979)

Therapeutic action
and method for mass production (1939-42)

Nobel Prize in Physiology and Medicine 1945

Of the 10 million soldiers killed in World War I,

about half died not from bombs or shrapnel or bullets
but rather from infection in an often relatively minor
Eric Lax in The Mold in Dr. Floreys Coat (2004)

The serendipitous discovery of penicillin (1928)

" Like many other bacteriologists, I have had many culture pla
with mould spores which drop in from the air, and like every ot
I have cast them out with suitable expressions of annoyance;
but on this one occasion I did not cast it out, and penicillin wa
It seems to me, as it must seem to you, most extraordinary
that a stray mould spore, coming from no one knows where,
settling in St. Mary's Hospital on a culture plate where it was n
should eventually have led to such phenomenal results."

Sir Alexander Fleming, Times, Londo

Therapeutic action

The Lancet (24 August 1940)

Therapeutic action
How to extract penicillin from its multi-substance broth
without rendering it impotent?
Eric Lax in The Mold in Dr. Floreys Coat (2004)

First animal assay: May 25, 1940

In one critical test, of eight mice infected with
streptococci,
four were given penicillin and four were not.
The four that had been treated Times,
lived:London
the (1975)
others died.

Penicillin: assays on mice

The Lancet (24 August 1940)

Penicillin: clinical trials

The Lancet (16 August 1941)

Could not be saved because not enough penicillin was available!

Penicillin production
at the Sir William Dunn School in Oxford
Counter-current distribution
apparatus designed by Heatley
for penicillin extraction
(the frame is from a discarded
bookshelf)
Total cost: ~ 5

Norman Heatley
(1911-2004)

Penicillin Girls

Penicillin production in the USA

On June 27, 1941, Florey and Heatley boarded a plane to the USA (via Lisbon)
On July 14, 1941, arrived at USDAs Northern Laboratory in Peoria (near Chicago)
Heathley stayed there for several months to work with Dr. Andrew Moyer,
improving fermentation procedures:
Changed culture conditions (use corn steep liquor)
Identified better producing strain
(Penicillium chrysogenum, from spoiled cantaloupe)
Florey convinced US drug companies to get involved
(Merck, Squibb, Pfizer)

Anne Miller (March 1942, New Haven)

First patient to be treated in the USA,

Suffered of blood poisoning, was given 5.5g
(at the time, this was half of the entire amount available in the
USA)
Recovered and died in 1999 at the age of 90

Penicillin use during WWII

Life magazine 1944

Structure of penicillin

Penicillin was far too small and simple a molecule to be a

Eric Lax in The Mold in Dr. Floreys Coat (2004)

The first time [Dorothy Hodgkin] heard of penicillin was whe

she accidentally met Chain in the street in Oxford in May, 194
He was very excited. He and Florey had just carried out their
on the antibacterial properties of Fleming's penicillin mould.
" Some day ", said Chain, "we'll have some crystals for you to

Times, London, April 21, 1

Dorothy Crowfoot Hodgkin (1910-1994)

Nobel Prize in Chemistry 1964

Structure of penicillin (1945)

Penicillin is a -lactam
Dorothy Hodgkin also solved the structure of cholesterol,
vitamin B12 and insulin

Basidiomycetes

Basidiomycetes (club fungi)

(~30,000 species)

Maiden veil fungus

(Dictyphora)
Fly agaric
(Amanita muscaria)

Basidium=latin for little pedestal

Diverged from Ascomycota ~900 million years ago
Asexual reproduction is rare
Basidiospores

Cryptococcus neoformans
a basidiomycetous yeast

Buchanan (1998)

Pathogenic in immuno-compromised patients (cryptococcosis)

Infection by inhalation
Encapsulated (polysaccharide capsule)
Budding
Genome sequenced in 2005; 19 Mb; 14 chromosomes; 6,572 genes

Symbionts
Mycorrhizae (plants)

Lichens (green algae or cyanobacteria)

Fungus-Animal symbiosis
Account for 1/5 of all known fungi

Ants carry back leaves to their nests,

so that fungi can break the leaves down
in a substance that the ants can digest

Fungus-gardening insects

Lichens
Shape and structure of the lichen depends on the fungal component
Ascocarp of fungus
Soredia
Fungal Algal
hyphae layer

Algal cell

10 m

Fungal hyphae