Culture of fungi

Saprotrophic fungi can be subcultured on media containing nutrients appropriate to

theirgrowth and development. Several different types of media have been used
successfully.The most commonly used in undergraduate classes consists of a fruit or
vegetable, ortheir extracts, mixed with sugars and agar, and set in Petri dishes. The
organic andmineral fractions are designed to supply nutrients similar to or commonly
found in theenvironment of the fungus. A few commonly used materials include:
•
Soil [SOIL AGAR]
•
Potato [PDA]
•
Tomato plus other vegetables [V8 JUICE Agar]
•
Malt extract [MA]
•
Dung [DUNG AGAR]
Approx 3 pellets of rabbit dung or equivalent is placed in 100ml water. agar is added, the
mixture autoclaved and plates poured. ..

These can be more highly defined by replacing the organic component with known
organic materials including:
•
Nutrient Dextrose [NDY]
•
Sabouraud dextrose [SABOURAUD AGAR]
Most of these media are available from commercial sources. Commercial sources
havea more consistent quality than those prepared from the raw materials each time. A
widerange of media are used for growing fungi.Most mycologists develop preferences
forcertain types of media based on experience and peculiarities of the type of fungi that
areroutinely grown.Media will affect colony morphology and color, whether
particularstructures are formed or not, and may affect whether the fungus will even grow
inculture.For example, some fungi lack the necessary enzymes to utilize different
carbonsources.All fungi require several specific elements for growth and
reproduction.Therequirements for growth are generally less stringent than for
sporulation, so it is oftennecessary to try several types of media when attempting to
identify a fungus in culture.Most fungi thrive on Potato Dextrose Agar (PDA), but this
can be too rich for many fungi,so that excessive mycelial growth is obtained at the
expense of sporulation.I havefound that most of the fungi isolated from soil, or from
substrates in the soil, i.e., plantdebris, grow well on Corn Meal Agar (CMA), a relatively
weak medium compared toPDA.Similarly, wood-inhabiting fungi and dematiaceous
(dark pigmented) fungi oftensporulate better on CMA or Oat Agar, both of which have
less easily digestiblecarbohydrate than PDA.Cellulose-destroying fungi and spoilage
fungi retain their abilityto produce cellulase when grown on a weak medium such as
Water Agar (WA) or PotatoCarrot Agar (PCA) with a piece of sterile filter paper, wheat
straw or lupin stem placed onthe agar surface.The introduction of pieces of tissue, such
as filter paper, wheat straw,rice, grains, leaves or dung, often produces good
sporulation dependent on theorganism grown.
Constituents of Media
Media generally contain a source of carbon, nitrogen and vitamins.Glucose (dextrose)is
the most widely utilizable carbon source, and hence is the most commonly used
ingrowth media.Fructose and mannose are the next most commonly utilized sugars by

fungi and are found in media from natural sources.Sucrose (table sugar) may be usedin
some media.Nitrogen sources include peptone, yeast extract, malt extract, aminoacids,
ammonium and nitrate compounds.Casamino Acids, a Difco product, is acid-hydrolized
casein, a mixture of amino acids. It is a good general source of nitrogen but isvitamin-
free.Bacto-Peptone, another Difco product, contains nitrogen and a highpeptone and
amino acid content. Salts, including Fe, Zn and Mn, are often added to‘defined’ media,
but are usually not added to the common media used for routineculture.fungi have
natural deficiencies for vitamins that are satisfied at mM to nMconcentrations.The most
common naturally occurring vitamin deficiencies are thiaminand biotin.Deficiency of
both is quite common among the Ascomycota.Other organicnutrients such as glucose
are often contaminated with vitamins sufficient to supply thegrowth requirements of
fungi.

Bacteria are suppressed by adding antibiotics to the agar. As most antibiotics

aredenatured by heat, the antibacterial agents are usually added to sterilised molten
agar,that is just above setting temperature (hand warm, but not hot to touch). Penicillin
(50units per ml), Streptomycin (50 unit per ml), Tetracyclin (30 units per ml) are
commonlyused this way, either alone or more commonly, in combination.
Chlomamphenicol (50mgper l) can be autoclaved and is added during preparation.