Protocol No. & Title: 3.1 Preparing a Pressed Sample of the Herbarium Version Date: 25February 13: Version 1 Author: Dr. Marsha J. Lewis and Dr. Tom Wendt Background and A herbarium (plural: herbaria) consists of preserved plant specimens, each with a label bearing documentary information. The primary use of the specimens is to study the variation and classification of plants, but they are also used as references for comparison and identification with unknown samples, documenting species distribution, identifying times of fruiting and flowering, and, in our case, documenting the specimen that we baited fungal cultures from. Purpose: The University of Texas Plant Resources Center (TEX-LL) has over 1,000,000 specimens and is the largest herbarium in the southwestern United States. The herbarium ranks fifth among U.S. university herbaria and tenth across the nation. Over 200,000 of its specimens are from Texas, which makes it the largest holding of Texas plants in the world. About one half of the specimens are from Latin America, with an especially strong representation of Mexico and northern Central America. Presently, the number of vascular plant collections inserted in the herbarium is growing at a rate of 6,000-12,000 specimens per year. This is an impressive resource that is literally outside our back door in the UT Tower! We will contribute to the herbarium collections by submitting the plants we select for baiting fungi. This way, we will have complete and thorough documentation of our plant tissue fungal species from plant to final organic products. This plant voucher pictured above was collected by Charles Darwin in September 1835 on the Galapagos Islands. Your voucher will live on in posterity! Materials: Safety Notes: 1/5  Plant press ends and stops  Cardboard 18”x12” with channels running the 12” way  Newspaper cut to size  Your plant specimen with stem, leaves, buds, seeds, etc. Bring a partner. 3.1 Preparing an Herbarium Sample TW (1) 1. Go to site where plant specimen is likely to be found and locate plant specimen. 2. Record the location as accurately as possible on the specimen bag with permanent marker. It is best to use a plastic bag and bring a wet paper towel to prevent the specimen from shriveling. If possible, include GPS coordinates. The goal is for others to find the location without you. 3. Using scissors or plant shears, cut the plant to include a leaf set and stem. We are not collecting root tissue at this time. The specimen should include at least the terminal parts of aerial shoots, including leaves and reproductive parts (flowers, seeds, fruits) if present. The plant sample should be representative (not the biggest or the prettiest!). Collect at least a couple of specimens of each plant. 4. Place the cut plant specimen in the labeled plastic bag. Label the paper specimen bag with plant name, time and date of specimen collection. Also, note anything of interest around the location. Be as detailed as possible. Do NOT rely on your memory! 5. Record a unique number to identify your specimen. Use your initials in front of the number to ensure the number is unique to you. You may start at “1” or “2013100”. It is up to you as long as it is recorded. 6. Upon return to lab, transcribe all information gathered about the plant and location into your lab notebook. Sketch the location and plant specimen if necessary. Also, write the unique number to identify your specimen in your lab notebook. 7. Prepare the herbarium sample as soon as possible. Some plants will wilt fairly quickly. NOTE: Store the plant parts you will use to harvest endophytic fungi in the 4 °C in a separate plastic bag. The plant should not be stored for longer than 72 hours for harvesting fungi. 8. The keys to achieving well pressed plants are to dry them as quickly as possible and to take care of the specimens as they dry. The following steps 2/5 3.1 Preparing an Herbarium Sample TW (1) describe making the plant press and Figure 1 (a and b) illustrate the plant press. a. Place a press cardboard on the work surface and open an appropriately sized folded single sheet of newspaper on top of it. Place the plant specimen on half of the open newspaper sheet. Arrange the plant specimen to show both underside and topside of leaves (i.e. all botanical features), and make sure that leaves are not covering critical features like flowers. You may need to trim a few leaves or branchlets off if there is a lot of overlap of parts. b. Make sure the specimens will fit on the mounting card when dry and leave space in the right hand corner for the label. The mounting card is 16.5” X 11.5”, smaller than a standard plant press cardboard. c. Fold over the newspaper on the top of the specimen, and write your collection number on the newspaper. d. Place a press cardboard on top of the newspaper, such that it is now sandwiched between two cardboards. Use corrugated cardboard to allow air to circulate through the press. e. Repeat the previous process until all specimens are pressed, then place the several plant/newspaper/cardboard sandwiches together and place wooden boards or lattices on the top and bottom. f. Around the stack, wrap belts, ropes or straps to hold the press together and apply even pressure. 3/5 3.1 Preparing an Herbarium Sample TW (1) Figure 1. a. (left) Illustration of plant press showing sequence of materials. b. (right) Photograph of plant press ready for the dryer. 9. Normally specimens take 3 to 7 days to dry completely in a dryer that has controlled humidity, air temperature, and air circulation. It’s much better to leave them too long than not long enough 10. After your specimen is dried (leaves with snap, not bend), your specimen needs to be mounted as follows: NOTE: Your specimen will actually be mounted by staff at the herbarium to maintain consistency between samples and ensure sample quality. You will be required to prepare the label for the mount. The label should be about 5(cm) x 10 (cm) and printed on archival (acid-free) paper, which the herbarium can provide. a. Obtain 16.5” X 11.5” mounting card b. Arrange and attach specimen with glue. Leave 1 cm border around the edge of the mounting card. Orientation of the plant should represent their habitat (e.g. flowers towards the top). c. Attach the specimen using mounting glue. d. After glue dries, sew if needed. e. The label contains the information about the specimen copied from the field notebook plus additional notes relating to the species name and identification. At minimum, your label should contain: 4/5 3.1 Preparing an Herbarium Sample TW (1) i. Family and scientific name ii. Collector’s name (s) and specimen number iii. Date of collection iv. Locality where plant was collected (state, county, locality, geographical coordinates if available) v. Habitat or type of plant community vi. Any other relevant details 5/5 3.1 Preparing an Herbarium Sample TW (1)