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MYCORRHIZAS FOR FORE.STRY AND AGRICULTURE. Chapter I Introduction
E. Recommendations
The presence of arbuscules should be used to identify VAM
associations and the presence of a Hartig net should be used to
define ECM associations. However, these definitions cannot
always be used and careful judgment may be required when
examining mycorrhizal associations in roots collected from the
field, particularly if roots are old, or associations are atypical.
Some older reports of the mycorrhizal status of hosts should be
disregarded if they do not include evidence that mycorrhizal
definitions were rigorously applied or anatomical investigations
were carefully conducted.
There is disagreement about whether arbuscular mycorrhizas
or vesicular-arbuscular mycorrhizas is the most appropriate name
for these associations (Section lA). It is important that scientists
use consistent terminology, because this influences the capacity of
computer searches to find papers, abstracting services to
categorise them, and the accessibility of mycorrhizal literature to
other scientists and students. It seems that the name arbuscular
mycorrhizas is gradually replacing vesicular-arbuscular
mycorrhizas in the scientific literature. It is best to use both the
words arbuscule and Glomalean in the title or keywords of
papers to ensure they will be retrieved by computerised search
programs in the future.
The terms ectomycorrhiza, ectomycorrhizas, ectomycorrhizal
should be used for ECM associations.
The degree of mycorrhizal formation of a host plant can be
expressed as the proportion (%) of root length colonised by
mycorrhizal fungi (see Chapter 4). However. roots with a
periderm (bark) layer resulting from secondary growth should be
excluded, since they have no cortex. In ecosystem surveys. it is
best to express the degree of mycorrhizal colonisation as the
proportion of roots available for colonisation that were
mycorrhizal. This requires an understanding of the dynamics of
root growth and mycorrhizal formation.
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MYCORRHIZAS FOR FORE.STRY AND AGRICULTURE.
A. Dissecting microscope
Eyepiece ~
fooIIl--- - - - - - - Objective
~I--_ _ _ _ _ _ _ Microscope stand
_ c_ _ Eyepiece
.......
__- - - Prism housi ng
Microscope
stand Revolving nosepiece
. .......
_ - - O bjective
Condenser
adjustment
...-------
...
Specime n ho lder
Stage
Condenser diap hragm
C o ndenser
Coarse
focusing
... Field ir is diap hragm
Microscope base
.....-- with light sou rce
Figure 1. 13. Mojor components of dissecting (A) ond compound (B) microscopes.
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MYCORRHIZAS FOR FORESTRY AND AGRICULTURE Chapter I Introduction
A. Dissecting microscopes
These microscopes normally have two eyepieces to provid e
stereoscopic vision and are most often used t o view whole
structures at relatively low magn ifications. In mycorrhizal studies
they are used to quantify mycorrhizal roots (Chapter 4), t o help
identify fungi (Chapters 2, 3), check fungal cultures (Chapter 5),
and to assess t he results of mycorrhizal inoculation (Chapter 6).
Some notes on dissecting mic roscope use are provided here.
I. It is advisable to keep all microscopes separate from areas where
samples are prepared to minimise exposure to dust and liquids. If
samples contained in liq uids (cleared and stained roots, etc.)
a re used ro uti nely, it is advisable to have a clear plastic
platform made to protect the base of the microscope.
2. Several types of illumination are required for mycorr hizal
studies.
(i) Incident-light illumination is req uired to required to co unt
spores of VAM fungi and select them to start pot cultures
(Chapter 3) and to view unstained roots, fungal fruit bodies,
etc. It is best to use a cool light source when observing living
material such as VAM fungus spores (halogen lam ps produce
less heat than incandescent light bu lbs of similar intensity).
Fibre-optic adjustable arms or ri ng lights mounted on the
objective lens are very useful (obtained from microscope
manufacturers) .
(ii) A transm itted light source, wh ich is built into t he microscope
base, is required to assess mycorrhizal colo nisation in cleared
and stained root samples.
(i ii) Darkfield illumination is not essential, but allows unstained
hyphae and roots to be observed more easily.
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MYCORRHIZAS FOR FORESTRY AND AGRICULTURE
B. Compound microscopes
A good quality compound microscope is an essential tool for
mycorrhizal research, to allow fungus identification by examining
fungal structures (Chapters 2, 3), and to examine sections of
mycorrhizal roots or mounted segments of cleared and stained
roots (Chapter 4). Microscopes should have powerful built-in
illumination, interchangeable objectives including a IOOX oil-
immersion objective and a fully adjustable condenser and dual
eyepieces. Some notes about microscope use are provided below.
I. All parts of the microscope must be kept free of dust and
cleaned carefully when they become dirty. Dust and dirt on
microscope components are the most common causes of
poor image quality.
2. Great care must be taken when examining liquid preparations
such as unsealed slides. fungi growing on culture media, etc.
to avoid contact with objectives. Immersion oil must be used
very carefully to avoid damage to other objectives. Objective
lenses are also easily damaged by abrasion.
3. The microscope condenser must be carefully adjusted so that
illumination of the specimen is uniform and sufficiently bright.
Condenser adjustments are often required when objective
lens magnification is changed. Kohler illumination is required
to obtain uniform lighting of the specimen with a compound
microscope. This can be achieved by raising the condenser
until the image of its iris diaphragm is in focus. centring that
image and then opening the iris beyond the field of view of
the objective that is being used.
4. Light microscopes may have additional components to al low
polarised light. interference contrast. phase contrast. o r
fluorescence microscopy (see Chapter 4). These may be of
great value in mycorrhizal studies, but require different
accessories to be installed and these parts are often very
expensive.
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MYCORRHIZAS FOR FORE.STRY AND AGRICULTURE. Chapter I Introduction
c. Photography
Microscope photographs are commonly used to document
taxonomic features of fungi and the results of experiments. A
relatively high quality microscope with a dedicated camera and
photomicroscopy controller is normally required to obtain
satisfactory results. Some information on microscope
photography is provided here. but more specific instructions are
provided in the manuals provided with photomicroscopes.
I. Photographic exposures are usually controlled by a dedicated
camera control unit. It is often wise to bracket exposures
(± I or 2 F stops).
2. The cleanliness of microscope parts. and proper adjustment
of the condenser (see above) is most important when
photographs are taken. as microscopy errors are often
magnified when slides or negatives are printed.
3. Colour photographs can be taken with a microscope by using
a film which is colour-balanced for the light source used
(usually tungsten). The light source voltage of the microscope
also needs to be adjusted to obtain the correct colour
temperature (typically 3200° K). It is normally best to use a
relatively 'slow' slide film (100-200 ASA) such as Ektachrome
EPY for microscope photography to obtain sufficient detail of
the subject material. but faster film is used for some
purposes. Fine-grained black-and-white negative film is also
commonly used for photography.
4. It is essential that records of information pertaining to
photographs be recorded. on a record sheet or log book.
when they are taken. Essential information includes the
exposure number. magnification. details of the subject
material. microscope procedures. or stains used. the date and
photographer's initials. This will allow photographs to be to
be identified and correctly labelled after processing.
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Chapter 2 Working with
MYCORRHIZAS FOR FORESTRY AND AGRICULTURE Ectomycorrhizal Fungi
Chapter 2
WORKING WITH
ECTOMYCORRHIZAL FUNGI
2.1. INTRODUCTION
This chapter concerns methods used for the collection,
description, management and identification of larger fungi
(Fig. 2. 1). Most emphasis is on mycorrhizal fungi, especially those
which associate with eucalypts in Australian forests. However,
these methods are also applicable to non-mycorrhizal fungi and
fungi which fruit on substrates other than soil. Terms used to
categorise fungi throughout this chapter are explained below.
Refer to Section 1.3 for an introduction to ECM associations.
A. Fungal taxonomy
Taxonomy provides a framework on which to base biological
work by providing names of organisms and by giving data about
the biology of the organism and its relationship to other
organisms. Communication between scientists relies largely upon
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MYCORRHIZAS FOR FORESTRY AND AGRICULTURE
"
Describing microscopic features
A. Observing spores and other structures ~ Section 2.4
B. Using microscopic information
"
Managing fungal collections
A. Fungal herbaria ---. Section 2.5
B. Database information
"
Identifying fungi
A. Distinguishing major groups ---. Section 2.6
B. Classification within major groups
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Chapter 2 Working with
MYCORRHIZAS FOR FORESTRY AND AGRICULTURE Ectomycorrhizal Fungi
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MYCORRHIZAS FOR FORE.STRY AND AGRICULTURE.
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Chopter 2 Working with
MYCORRHIZAS FOR FORESTRY AND AGRICULTURE Ectomycorrhizol Fungi
Table 2.1. Ectomycorrhizal an d ectendomycorrhizal fungal taxa. List compiled from data in Miller 1982,
= =
Ken drick 1992, Molina et al. 1992 an d Bougher 1995. (H hypogeous truffle-like fungi, E epigeous fu ngi,
P = puffballs, genera w ith an aste r isk have many non-mycorrhizal species, genera in bold are known to occu r
in Australia.)
Family Genera Type
HYPHOMYCETES
Cenococcum, ete. Co ni dia l
ZYGOMYC ETES
Endogonaceae Endogone, Sclerogone H
ASCOMYCETES
Ascobolaceae Sphoerosomo H
Balsami aceae Bo/somio, Picoa H
Elaphomycetaceae E/ophomyces H
Geneaceae Geneo, Geneobo H
Geoglossaceae Geoglossum*, Leotio*, Trichoglossum E
Helvellaceae Gyromitro, Helvello E
Barssia, Fischerula, Gymnohydnotryo, Hydnotryo, Mycoclelondio H
Pezizaceae Aleurio*, Pezizo*, Phillipsio*, Pulvinio, Sarcosphaera E
Amyloscus, Hydnotryopsis, Mucitu rbo, Pochyphloeus, Pezizo, Ruhlondiello H
Pyro nemataceae Geopora, Humaria, Jo fneodelphus*, Lomprosporo* Sphoerosporello,
Trichoph aea, Wilcoxina E
Choiromyces, Dingleyo, E/derio, Geopora, Hydnobolites, Hydnocystis,
Lobyrinthomyces, Pourocotyis Reddellomyces, Sphoerozone, Stephensia H
Sarcoscyphaceae Plectonio*, Pseudoplectonio*, Sorcocypho* E
Te rfeziaceae Choiromyces, Terfezia H
Tuberaceae Mukogomyces, Paradoxa, Tube r H
BASIDIOMYCETES
Amanitaceae Amonito, Limocello E
Torrendio H
Astraeaceae Astraeus E
Pyrenogaster, Rodiigero H
Boletaceae Austroboletus, Boletellus*, Bo/etochoete, Boletus, Buchwoldoboletus*, E
Cho/Ciporus, Fistulinello, Gyrodon, Gyroporus, Heimiello, Leccinum, Phlebopus,
Phylloporus, Pulveroboletus, Rubinoboletus, Suillus, Tylopilus, Xonthoconium
Alpovo, Boughero, Ch omonixio, Gostroboletus, Rhizopogon, Royoungio H
Truncocolumella H
Camharellaceae Conthorellus, Croterellus E
Chondrogastraceae Chondrogoster H
Clavariaceae Aphelorio, Clovorio, Clovoriodelph us, Clovicorono, Clovulino, Clovulinopsis,
Romorio, Romoriopsis E
Cortici aceae Amphinema, Byssocorticium, Byssosporia, Piloderma, ete. E
Cortinariaceae Astrosporino, Cortinorius, Cuphocybe, Dermocybe*, Descoleo, Hebelomo, E
Inocybe, Leucocortinarius, Rozites, Stephonopus, etc.
Cortinorius, Cortinomyces, Descomyces, Destuntzia, Hymenogoster, H
Quodrisporo, Setchelliogoster, Thoxterogoster, Tim groveo
Cribbiaceae Cribbeo, Mycolevis H
Entolo mataceae Clitopilus, Entolomo, Leptonio, Rhodocybe* E
Rhodogoster, Richoniello H
Elasmomycetaceae E/osmomyces, Gymnomyces, Mortellio, Zelleromyces H
Ge lopellidaceae Gelopellis H
Go mphaceae ~~m E
Gom phidiaceae Chroogomphus, Cystogomphus, Gomphidius E
Hydnaceae Bankera, Dentinum, Hydnellum, Hydnum, Phellodon E
Hygroph oraceae Bertrondio* Com orophyllus, Gliophorus, Humidicutis*, Hygrocybe,
Hygrophorus*, ete. E
Hysterangiaceae Hysterongium, Pseudohysterongium, Troppeo H
Leucogastraceae Leucogoster, Leucoph/eps H
Lycoperdaceae Lycoperdon* p
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MYCORRHIZAS FOR FORE.STRY AND AGRICULTURE.
Melanogastraceae Melonogoster H
Mesophelliaceae Costoreum, Diplodermo, Gummiglobus, Molojczukio, Mesophellio,
Nothocostoreum
Octavianinaceae Octovionino, Sclerogoster H
Paxillaceae Poxillus E
Pisolithaceae Pisolithus P
Polyporaceae Albatrel/us E
Russulaceae Loctorius, Russulo E
Archongeliello, Cystongium, Mocowonites H
Sclerodermataceae Sclerodermo P
Horokiello, Sclerodermo H
Sedeculaceae Sedecula H
Stephanosporaceae Stephanospora H
Strobilomycetaceae Strobilomyces E
AU5trogautieria, Chamonixia, Gautieria, Wakef/eldia H
Thelephoraceae Boletopsis, Thelephoro E
Tricholomataceae Clitocybe*, Cystodermo*, Conthorellulo, Catathelasma, Loccorio, Lepisto, E
Leucopoxillus, Tricholomo, Tricholomopsis*
Gigasperma, Hydnongium, Podohydnongium H
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Chapter 2 Working with
MYCORRHIZAS FOR FORESTRY AND AGRICULTURE Ectomycorrhizal Fungi
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MYCORRHIZAS FOR FORESTRY AND AGRICULTURE
Cover t
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