4. Fungi

4.1 Voucher Requirements

Diversity Inventory

Collect and process one of each type of macrofungal fruitbodies within a study area.

4.2 Data Needs

The Collaborator Collection Slip should be completed and submitted with the specimen (Figure 4). If possible, a literature reference for the species description should be included in the space provided, if the identifier is not a taxonomic authority, or if the reference is obscure. Additional field notes (space at the bottom of the slip, and additional data, sketches, or photographs may be submitted along with the form) are essential for agarics and other fleshy fungi whose appearances change dramatically after drying. Temporary color changes to fleshy fungi after bruising or wounding should also be noted, as these often disappear after heat treatment. Spore prints of agarics should also be included. Habitat notes are also useful.

The following is the minimum voucher data that must be included:

• Field Collection Number: A unique field label should be assigned to each specimen. A lifetime system is recommended rather than labels for each project or year. A label may look like "Shepard 2094" where the collector and number are unambiguous.
• Collection Date: Format for dates is YYYY/MM/DD. Note that four digits are used for the year.
• Detailed Location: It is important that the place of collection can be precisely located in the future. Include country, province, city, and gazetted location name. Location must include latitude and longitude or Universal Transverse Mercator (UTM) coordinates (use NAD83 when using a GPS or a 1:50,000 map), as precisely as can be calculated, plus a brief written description, noting elevation, as well as direction and distance from a conspicuous landmark.
• Recommended Additional Data: Many aspects of the specimen and its environment may be useful to record. Check with curators for suggestions to enhance the value of specimens.
• Collector name: It is important to note the collector's name. This acknowledges this person and is useful when details need to be tracked should data be misplaced or additional information desired.
• Habitat: Basic descriptions of the collection area should be made in as much detail as is practical. Aspects that may be useful include vegetation, associated flora and fauna, altitude, cover, depth of collection, substrate, pH, salinity, nutrient load, and light conditions.
• Collection method: The method used to collect the specimen.
• Mode of Acquisition: How the museum acquired each specimen (gift, etc.).

Figure 5. Collaborator collection slip.

4.3 Preparation and Care of Specimens

4.3.1 Photo Documentation

For photography consider the following: tripod, flash, shutter release, reflectors, umbrella (to protect the camera), water spray bottle (to freshen or clean specimens in situ), scales (mm and cm). See also section 2.3.1.

4.3.2 Whole Specimens

For more complete collecting and preserving procedures see Callan (1998) and the species manual by RIC (1997), Inventory Methods for Macrofungi (No. 41) sections entitled Field Equipment, Field Procedures, Processing Spores, Recording Data on Fresh Specimens, and Preserving Herbarium Specimens.

Preparation and submission specifics given below can be found on the Pacific Forestry Centre's website at http://www.pfc.cfs.nrcan.gc.ca/. Its Forest Pathology Herbarium abbreviation is DAVFP (Department of Agriculture, Victoria, Forest Pathology). DAVFP information is under the heading "Forest Biodiversity". You can contact the Forest Pathology Herbarium's curator Dr. Brenda E. Callan or the technician Ms. Analie Fernando by phone at (250) 363-0684.

General preparation protocol to meet criteria for specimen accession can be found in section 4.4. Below fungi collection protocols are provided according to type of specimen.

A) Foliar Disease:

• Cut one or more lengths of branch or stem, ca. 20 cm long, with diseased foliage attached. Include a second cutting with healthy foliage, for comparison.
• Include flowers or fruit if identity of plant is unknown.
• If collection is made in spring or early summer, pathogen fruiting bodies on current year growth may be immature. Search for overwintered foliage in ground litter or branch crotches, and include some, either pressed or wrapped in a paper bag, with the fresh sample.
• Do not moisten specimen. This encourages growth of contaminants.
• Do not seal specimen in plastic bags or wrap. This encourages growth of contaminants. It is better to let the material dry.
• Press the freshly-cut specimen between folded sheets of dry newspaper. If specimens are too dense and bushy to press, place in bottom of large paper bag, fold bag over, and tape shut.

B) Stem or branch canker (<10 cm diameter):

• Sample should include both living and dead host tissue, if possible, because the pathogen is most active in dying tissue (i.e. be sure to cut stem or branch a few centimeters below the visible edge of the canker).
• Saw or clip a ca. 20 cm long section from a cankered area of branch or stem.
• Pack sample in newspaper (not plastic), or roll it inside a large paper bag.

C) Stem or branch canker (>10 cm diameter):

• If tree is to be felled, or if it is possible to remove a large branch: cut a 10-20 cm long section of canker, preferably at the lower edge of the canker, where there is the greatest likelihood of living tissue.
• If tree cannot be felled, use a hammer and chisel, knife, or axe to cut out a ca. 10x10 cm square section of bark from the edge of the canker.
• If fruiting bodies are visible on other parts of the canker, remove a similar-sized portion from this area, too.
• Wrap sample in newspaper (not plastic).

D) Wood decay samples:

• Look for conks, mushrooms or other fruiting bodies which might be associated with the decay.
• Break off, or cut out conk, wrap in newspaper, and if it is not fragile, include in package with decay sample. If mushrooms are observed, collect and process as described below in Section G.
• Do not include large pieces of substrate in the same bag with fragile fruiting bodies. Large clumps of substrate either soil the fruiting bodies, or smashes the fungus to pieces in transit.
• Document the type of wood decay (brown cubical, white rot, laminated decay, white pocket rot).
• To collect a sample of wood decay, cut into wood near the conk or point of breakage, using an axe or chainsaw. Ideally, wood sample should contain symptoms of advanced decay, incipient decay, and sound wood. These conditions are found near the edge of the decay column. Wood sample should be at least 15 x 15 x 15 cm; large enough to split with an axe.
• Do not send in increment cores, or small chips. These are too small to culture or identify. Avoid areas with insect activity, and weathered, exposed wood. Samples made from these areas are contaminated with bacteria and molds.
• Wrap sample in newspaper (not plastic).

E) Standing dead or dying tree with suspected root disease:

• Sample from a dying or recently dead tree in the stand. These trees are often found in the middle of a root disease center, with old dead trees in the centre, and symptomless trees at the edge.
• Samples should be made from ground-level areas of the butt of the tree where there are visible signs (mushrooms, conks) or visible symptoms (heavy pitch exudation, cracked bark at butt of tree). Use an axe, polaski, or chain saw, and proper safety techniques, either falling the tree or removing from a standing tree a chunk of the outer part of the butt, 20 x 20 x 10 cm, important: leave bark intact on sample.
• If no root disease symptoms or signs are apparent on the butt of the tree, and examination of above-ground cambium reveals no evidence of root diseases (stains, decay, or mycelial fans), excavate the roots at the base of the tree, looking for pitching, stains, decay, or mycelial fans, and obtain a sample as described above. If the root is <10 cm diameter, cut out a 20 cm long, whole section of root, bark intact.
• Wrap sample in newspaper (not plastic).
• NOTE: Long-dead, standing trees are generally colonized with saprophytic decay fungi. These fungi often produce mycelial fans or decay columns which extend several meters up the trunk of a dead tree, and are sometimes confused with pathogenic fungi. Care should be taken to excavate down the butt and along roots of the tree, until signs of pathogen activity, such as pitching, or stunted, callused roots, are encountered. Samples should be taken from these areas.

F) Blowdown with decay - how and where to sample:

• Try to determine host species, at least to hardwood vs. conifer. If the tree is too old to identify, make note of other tree species in the vicinity.
• Note: fungi fruiting on dead, long-down trees are often not the same ones responsible for tree failure.
• If stem has failed, cut into wood near conk or point of breakage. Sample wood as described in Section D.
• If roots have failed, check for root decay and sample from decayed roots, either those remaining in the ground or larger roots in the exposed root plate (See Section E).
• Wrap sample in newspaper (not plastic).

G) Fungi fruiting on soil or duff:

• Dig the fruiting body out of the substrate with a knife. Make sure that the entire fruiting body is collected, as some fungi have large, below-ground structures.
• If the mushroom is on wood, also follow instructions for collecting a wood decay sample (See Section D).
• If possible, collect several fruiting bodies, especially if a number of them are at different levels of maturity.
• Temporary in-field packaging. These fungi are fragile and must be individually wrapped or packaged for transportation. Pack the fungus in a paper bag (never plastic) or, wrap in wax paper. Roll the wax paper in a cylinder around the mushroom, then twist each end like you would a candy wrapper. Because waxed paper tends to collapse in pouring rain and smaller specimens either become crushed by larger or heavier specimens, or water logged, aluminum foil is now preferred in temperate climates. Aluminum foil forms a ridged protectant, retains moisture, protects specimens from heavy rain, and does not stick to viscid pilei.
• Carry mushrooms in a rigid container so that they do not get squashed. For small fleshy fungi (mushrooms, cup fungi, coral fungi) small plastic tackle boxes with many compartments, or pill vials or film canisters may be used. For bulk collecting a reinforced backpack or carrying basket is recommended.
• Process the specimens the same day they are collected, using the following guideline:
• Obtain a Spore Print. Mushroom identifications depend on spore colour. A spore print is very helpful, but may only be obtained while the specimen is very fresh. To make a spore print, cut the stalk off a mature mushroom and place the cap gill-side down on a piece of paper. If you suspect the spores might be white (check the stalk or the ground underneath), use colored paper. Cover the cap with a drinking glass or bowl if it is a very dry day. Consider making the spore print outside if the mushroom was collected on a cold day; as it might stop sporulating when warmed up to room temperature inside. Spore prints appear in several hours or overnight. Protect the spore print between cardboard before shipping.
• Air-Dry, or Heat-Dry the Sample. Most unpreserved fungi turn to mush during the few days it takes to send a sample by mail to an expert. Specimens should be dried before shipping. Small, fragile specimens under dry conditions may be air-dried in opened paper bags or wax paper in a warm, dry room for a day or two. Large fungi should be quickly dried using a portable dehydrator (such as those sold for drying food), otherwise they will decay and/or become infested with maggots As a last resort, with careful and constant monitoring, a conventional oven may be used, if turned to the lowest setting with specimens on the highest possible rack with the door open. Alternatively, use the warming drawer below the oven, left partially open while the oven is in use. Ideally, drying temperatures should be around 50 C.
• Pack dried specimens in rigid containers, or in zip-locked plastic bags packed in a box filled with foam chips. Dried specimens are very fragile.