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Macrofungi
We define macrofungi to encompass those fungi that produce large, easily observed and collected sporocarps.
This includes the Agaricales in the broad sense (mushrooms and relatives), Aphyllophorales (polypores, tooth fungi, coral fungi, etc.), gasteromycetes (puffballs, etc.), and some groups of Ascomycetes, primarily discomycetes (cup fungi) Xylariaceae, and the genus Cordyceps.
Ecologically, these fungi include: (a) beneficial mutualists such as the many mushrooms and related fungi that associate to form ectomycorrhizas with species of Quercus and Alnus, (b) important plant pathogens such as species of Armillaria and some polypores, and (c) the dominant decomposers of forest systems which are vitally important in nutrient cycling.
In Asia, many of these fungi are reported to have medicinal uses including anti-cancer effects. These fungi have not been included rigorously in drug discovery programs or other bioprospecting studies outside of Asia. Protocols: Techniques for describing and processing macrofungi are given in "Guía para la Recolección, Descripción y Preservación de Macrohongos" (Milagro Mata, INBio, unpublished). Three types of sampling protocols will be employed: (A) random sampling, (B) sampling of fixed sized plots, and (C) sampling of a fixed number of downed logs. A) Random sampling Opportunistic collecting will be undertaken at 3 sites in each conservation area throughout the year.
B) sampling of fixed sized plots 1. One site within each Conservation Area will be selected to establish the permanent plot. The site will be representative of an important forest type in the Conservation Area, and will be chosen to optimize the diversity of forest types sampled within the country. The plot will be established within the site in an area as homogeneous as possible, as level as possible, and as easily accessible from the road as possible as it will be visited repeatedly. 2. The plot will consist of ten 100 meter long transects. Typically transects will be laid out parallel to one another with 10 meters between them. However, if the shape of the area to be sampled does not allow this, some of the transects can be laid out end to end. Transects will be marked by placing a flag or stake every 5 meters. Each transect will be assigned a unique letter, and each flag will be numbered sequentially within a transect (i.e.., A1-A20, B1-B20,....J1-J20). 3. Each person sampling will have a plastic pipe of wooden pole that is 1.262 meters long. They will move down each transect from one flag to the next using this pole to circumscribe 5m2 circular subplots. This gives twenty 5m2 subplots per transect, for a total sampling area of 1000m2 (0.1ha) per site. 4. All macrofungi occurring in a subplot will be collected, labeled with the transect letter and subplot number, and placed in an appropriate bag or container for transport back to the field station at the end of the collecting day for sorting, taking of descriptions, photographing, and drying. The substrate for each specimen (soil, leaf litter, wood) will be noted. Care will be used to not walk in or unnecessarily disturb the subplots. The macrofungi transects will be sampled 3-4 times per year, for 3 years. June (3-4 weeks after the start of the rainy season) August (the middle of the rainy season) November (3-4 weeks before the typical end of the rainy season) March (the middle of the dry season)
Restricting quantitative sampling to the 0.1 ha plots would exclude most of the fungi found on larger pieces of wood, as the frequency of these larger substrates within the plots would be very low. Therefore, this substrate must be sampled separately. The following protocols should capture a good percentage of the diversity, as well as provide quantitative data to enable some analysis of abundance and host and size class specificity. Logs will be chosen and marked at one site in each conservation area during 2000 and early 2001. Then they will be sampled 4 times per year (March, July, September and December) starting in year 2001 for 3 years.
1. Logs to be included in the sampling will be: 2. There will be approximately 30 logs in each of the following decay classes. Class 1: Relatively newly fallen, usually with its bark on. Class 2: Medium rotten, bark fallen off, and knife can penetrate 1-2 cm into the wood without undue pressure. Class 3. Thoroughly rotten, knife can penetrate into the log without much pressure; the wood can be partly destroyed with the fingers. 3. Each log will be marked with a colored plastic band and given a number. A map will be made so that the collector will see that she/he has missed a log if the numbers jumps from 27 to 30 without any intervening log. The length and diameter of each log will be measured and recorded. If the diameter of the log varies greatly from one end to the other, measure the diameter near both ends and at the middle. 4. In general, only 2-3 specimens of each species will be collected from each log. If there are many specimens of a particular species, a pin or other mark will be used to remind one not to recollect the same species on the next trip. 5. Small specimens such as corticoid fungi and ascomycetes will be collected whole; while large polypores will be sectioned radially in a width of approximately 1-1.5 cm to make drying quicker so to avoid mold. 6. The genus or species of the host tree will be noted when possible. Protocols for documenting and preserving fungi in the field: 1. Collection data for all specimens will be entered into a database on laptop computers. This database will be compatible with ATTA and will enable ready transfer to that web accessible system.
2. Macrofungi will be described and photographed before drying. The collections then will be warm-air dried and sealed in plastic bags for protection against breakage and high humidity that will cause mold growth before being transported to the laboratory.
Protocols for identifying and curating the collections: Identification of macrofungi is based both on macro- and micromorphological characters. Curators and technicians will use compound and stereo microscopes equipped with digital image capture systems to facilitate capturing images for documentation, rapidly sharing information with international collaborators through e-mail, publications and other products. Plot based inventories generate an incredible number of specimens, and an accession policy is mandatory to prevent a herbarium from being overwhelmed (both in processing time and space) by redundant collections. In general, it is recommended that 2 or 3 good samples of each taxon from each site be retained.
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