uid=ARC,o=lter,dc=ecoinformatics,dc=org all public read 2012_GS_ITEX_PF_ShootHarvestData.05 Arctic LTER Program Gaius Shaver

Ecosystems Center at the Marine Biological Laboratory 7 MBL Street Woods Hole MA 02543 United States

Arctic LTER Program

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7496 arc_im@mbl.edu http://ecosystems.mbl.edu/ARC/ Edward Rastetter

Ecosystems Center at the Marine Biological Laboratory 7 MBL Street Woods Hole MA 02543 United States

Associated Researcher Mathew Williams

University of Edinburgh School of Geosciences Darwin Building Edinburgh EH9 EJU United Kingdom

Associated Researcher James Laundre

Ecosystems Center at the Marine Biological Laboratory 7 MBL Street Woods Hole MA 02543 United States

Data Manager Laura van der Pol

Ecosystems Center at the Marine Biological Laboratory 7 MBL Street Woods Hole MA 02543 United States

Field Crew 2012 Leaf and plant tissue area and mass from shoots harvested from 19 1m x 1m point frame plots near Toolik Field Station, AK during the summer of 2012. Six shoots were harvested from each plot, two from each canopy layer: upper, middle, and low. Each shoot came from a different plant, and the species selected was based on the species dominant in that canopy layer. The leaf area and mass were used to correct A/Ci and light response curves taken on each shoot [data published separately]. At the time of collection, the location relative to the point frame, height, and leaf area index (LAI) of each shoot was measured; those data are included here. shoot photosynthesis shrub canopy point frame A/Ci curve light response curve A/Ci Curve leaf mass per area (LMA) leaf area index (LAI) ImageJ opaque conifer chamber Li-Cor 6400 Acceptance and utilization of LTER data requires that: The Principal Investigator be sent a notice stating reasons for acquiring any data and a description of the publication intentions. The Principal Investigator of the data set be sent a copy of the report or manuscript prior to submission and be adequately cited in any resultant publications A copy of any resultant publications should be sent to: Principal Investigator Ecosystems Center Marine Biological Laboratory Woods Hole, MA 02543 http://ecosystems.mbl.edu/ARC/meta_template.php?FileName=./terrest/tracegas/xlsfiles/2012_GS_ITEX_PF_ShootHarvestData.html Upland site; co-located in Block 1 of the Shrub LTER sites; IVO 68° 38'18.8" N, 149° 34' 07.2" W +/- 50m. Except for plots marked "FERT", plots are outside of the designated LTER treatments, though are exposed to the same environmental conditions. All plots were chosen by the dominant shrub canopy (either Salix pulchraor Betula nana) and preferentially selected to be 90cm+ in height. -149.568666666666 -149.568666666666 68.6385555555555 68.6385555555555 Outlet site; co-located in Block 2 of the Shrub LTER sites; IVO 68° 38'008.1" N, 149° 35' 017.1" W +/- 50m. Except for plots marked "FERT", plots are outside of the designated LTER treatments, though are exposed to the same environmental conditions. All plots were chosen by the dominant shrub canopy (either Salix pulchraor Betula nana) and preferentially selected to be 90cm+ in height. -149.588083333333 -149.588083333333 68.6355833333333 68.6355833333333 2012-06-23 2012-08-07 Organisms Studied Genus Betula Species nana Genus Salix Species pulchra Genus Salix Species glauca This was a season-long project, though it followed similar methods to ITEX projects performed starting in 2003 that are likely to be replicated in the future for reasearch at the Toolik Field Station, AK. Version 2: Missing values changed to #N/A. CH 28Jan2013 Version 3: Metadata updated to newer form (with sites sheet). CH April 2013. Version 4: Corrected eml excel file name wrong extension. JimL 16May13 Version 5: Change excel extension from xlsm to xlsx Jim L Data Manager

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7496 arc_im@mbl.edu http://ecosystems.mbl.edu/ARC/ ARC LTER

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7496 arc_im@mbl.edu http://ecosystems.mbl.edu/ARC/ HARVEST METHOD: The methods for setting up each point frame plot are described below in the section 'POINT FRAME PIN-DROP METHODS'. The methods here describe how each shoot was harvested from the plots which were used for the point frame method. Six shoots were harvested from each of 19 1m x 1m point frame plots which were dominated either by Salix pulchra or Betula nana tall, shrub species. The "shoots" described here are branch clippings between eight to ten inches long. Whenever possible they were selected for appearing relatively healthy, intact leaves, and each shoot was taken from a different plant. As these shoots were used for shoot-level and leaf-level measurements, shoots with bi-furcated stems -- or two stems from the same branch/height were cut -- one for leaf-level and the second for shoot-level analyses. Before being cut, we measured the distance from the point frame to the highest tip on the shoot as well as the distance from the point frame to the shoot five inches from the tip. This way, we could approximate the angle of the shoot relative to the ground. Using the same criteria as for the pin-drop measurements, we measured the distance from the top of the shoot to the soil. In addition, the row number and pin-hole number nearest to the shoot's location with respect to the point frame was also recorded . We then measured the leaf area index (LAI) of the shoot by holding an LAI-2000 (Li-Cor Inc., Lincoln, Nebraska, USA) in the exact location where the shoot had been and taking the average of three readings (one above, three below). For these measurements we used the one-quarter cut out and took care to always hold the instrument level and with the technician's body casting a uniform shadow over the instrument's eye, with the technician standing between the instrument and the sun. On occasion when it was raining during the shoot harvest, the area near where the shoot had been was marked with flagging tape, and the the LAI measurement was taken at a later date, using the height from frame/distance from ground meausrements as well as the row number and pin hole measurements as a guide. Once cut, the shoot was placed immediately into water and transported to the lab. Once in the lab, the end of each shoot was clipped under water to ensure that there were no air bubbles in the stem that would inhibit the flow of water. Shoots were then allowed to sit at ambient room conditions (~20-25 degrees Celsius) until the the photosynthetic rates could be measured. AREA CALCULATIONS: Area calculations were made using the open source software Image J, available at http://rsbweb.nih.gov/ij/download.html. The procedures for each kind of area calculation are outlined here: SHOOT SILHOUETTE AREA: Digital image capture of intact stem and leaves 1. Arrange the shoot on a white background (e.g. blank paper or foam board) as it sat in the LiCor conifer chamber. Add a note of sample number and date. 2. Place a ruler that you can later use as a reference. Ensure ruler/stem do not overlap. 3. Use a tripod to hold the camera in a steady position and as close as possible to the stem, keep this distance constant throughout the samples if possible to avoid the need to re-calibrate Image J. 4. Take a digital image of the shoot in very bright light conditions, using diffuser or reflector panels as needed to minimize shadows. [Shadows can be mistakenly analyzed in the software as plant tissue if not careful.] 5. Download the images in the computer, save them as TIFF files. SHOOT TISSUE AREA: Image capture of shoot tissues (leaf, petiole, stipule, organ, green/brown stem) using a scanner 1. Pluck the sample with forceps, arrange leaves, petioles, stipules, green and brown stems and inflorescences separately, grouped by type onto the white background of a plastic board. Place a ruler that you can later use as a reference. Add a note of sample number and date. 2. Press flat with the clear plastic cover. Ensure ruler/plant parts are not overlapping. 3. Place plastic side down on scanner (ensure no parts have overlapped when inverting plastic board). 4. Scan the images to the computer, save them as TIFF files. BOTH SILHOUETTE AND TISSUE AREA ANALYSES: In the Image J software program: 1. Open leaf image via Select File ? Open? your file name 2. Convert scanned colour image of leaf to greyscale: 3. Image?Type?8 bit 4. Set measurement scale: Draw a line over a 50 mm section of the ruler then Analyze ?Set Scale 5. In Set Scale window enter 50 into the 'Known Distance' box and change the 'Unit of Measurement' box to mm , check 'Global' (the Global option will apply the same scale to all the images you open after this, if you took the photos from different heights you might need to repeat this procedure every time) 6. Draw a new line and confirm that the measurement scale is correct. 7. Threshold the leaf image: Image? adjust? threshold 8. Play with sliders to include all of leaf in red and click ‘Apply’ 9. Calculate area of entire silhouette area and area of each tissue type: Enclose the region of the shoot or tissues that you wish to calculate the area for with the rectangular selection tool, then Analyze ? Analyze Particles 10. Select the options of display masks and display results. Area of measured pixels is displayed. Check that only your leaf is included in the outline (not the ruler or extraneous tissue types). The data of the leaf area is displayed on a new window ‘summary’, this data can be copied into Excel or the data can be saved directly as an Excel file. Keep summary window open between picture analyses to maintain a record of measured areas. 11. Save new black and white image as well as its original colour photo with an appropriate file name. DRYING AND WEIGHING: Once digital images of each shoot and tissue had been made, each plant tissue was separated, counted, and placed in a labeled coin envelope. These samples were then dried for a minimum of three days at 60 degrees Celsius prior to being weighed on a four-point balance with glass enclosure. Plant tissues were weighed in aggregate, thus the average mass included in these data are the total tissue mass divided by the count of that tissue (e.g. number of leaves, petioles, etc). In the same manner, plant tissure area was calculated in aggregate, thus the average area per leaf/petiole/stipule is based on the total area of that tissue divided by the count of that tissue. At the start of this project, we did not take care to distinguish leaves, petioles, or stipules. While not thought to make a significant difference in the area or mass calculations for most samples, we did not want to include the petioles in the CHN analyses. Thus, for those samples that had removeable petioles (all B. nana, most S. pulchra, but no Vaccinium spp. or L. palustre samples), the petioles were clipped after drying and the leaves re-weighed. In some cases the mass of the leaves with petioles removed was greater than the original mass; in these cases, the original mass was preserved. [The original mass of all samples weighed is listed in the ORIGINAL LEAF MASS W/ PETIOLE data-column.] NOTE: The area used for correcting the LiCor A/Ci and light curve data included leaves, petioles, and stipules. CHN ANALYSIS: Grinding: All leaf samples were dried in an oven at 60°C before grinding. Samples were small enough to grind the entire sample without subsampling. Leaves were ground using the Retsch MM 200 for 3 minutes or until a talcum powder consistency was achieved. Weighing: After grinding, samples were stored in glass scintillation vials and dried again at 60°C for at least 36 hours. Once samples were removed, vials were tightly re-capped. When not in use, vials were stored in dessicators. 3.5-4.5 mg of each sample was weighed into a 10x12 mm tin capsule. A standard calibration curve was created using increasing amounts of aspartic acid (from about 0.2 mg to 5.0 mg). A chemical standard, acetanilide, and an organic sample, apple leaf, were run after the standard curve. Every ten samples, an aspartic acid check standard and a duplicate of an already-packed sample were run. CN analysis: CN analysis was run between 10/4/2012 and 11/15/2012 by Rachel Rubin using the ThermoScientific 2000 at the Ecosystems Center, MBL, Woods Hole, MA. Duplicate sample values were averaged (mass, %N and %C) before inclusion into final results. The CHN data is available in the file "2012_GS_ITEX_PF_CHN_Data" POINT FRAME PIN-DROP METHODS: We preferentially selected tall shrub canopies dominated either by Betula nana or Salix pulchra, that is canopies that were greater than 75 cm height. Care was taken to select fairly uniform canopies, that is avoiding the edge of a shrub stand or areas where the canopy had a large gaps, suggesting the area may have been disturbed. We used point frames constructed from a 1.1 m x 1.1 m aluminum square with holes in each corner to accomodate steel rod posts used as the legs of the point frame. In this way, the frame could rest upon the four leg posts that had been hammered into the ground and remain adjustable in each corner. The frame had a level on each side, and great care was taken to ensure that the frame was (a) unable to be pushed deeper into the ground and, (b) level on all four sides prior to taking measurements. These factors were important to the measurement to have accurate data regarding the distance from the frame and the overall height of each point sampled in the canopy. The aluminum frame had numbered, regularly spaced holes on two opposite sides in order to accomodate a metal bar that could be placed across the frame and locked into place. [These holes on the frame are the row numbers.] The bar that was placed across the frame similarly had numbered, evenly spaced holes in order to accomodate a pin--a long (100-200cm) metal rod with a diameter of ~3.175 mm. [The holes on this bar are the pin hole numbers.] Measurements were only ever taken from odd row numbers, and alternated even/odd pin hole numbers with each row; in this way, for every plot 25 evenly spaced locations were sampled covering an area of one square meter. The length of the pin was marked every half-centimeter so that the distance could be read easily. Measurements were made by lowering the pin through a pin hole and, once encountering a leaf or stem, recording the following: row#, pin hole#, hit#, and the species hit. If the object hit was not a leaf, the plant tissue was noted; the diameter of each stem hit was estimated in millimeters, and the length of every graminoid blade hit was recorded from the point at which it was hit to the tip. As the primary species of interest for this project were for a select number of species (B. nana, S. pulchra, S. glauca, S. reticulata, V. uliginosum, V. vitis, L. palustre), species that were not the target of interest were classified as functional groups--e.g. graminoid spp., forb, moss. The last pin-hit recorded for each pin hole was always at the "soil" which was considered to be the transition between the green and brown plant material, often in a mossy layer. pers-1 http://www.blackwellpublishing.com/plantsci/pcecalculation/ Sharkey, T.D., Bernacchi, C. J., Farquar, G.D. and Singaas, E.L., 2007. Fitting photosynthetic carbon dioxide response curves for C3 leaves. Plant, Cell and Environment, 30: 1035–1040. doi: 10.1111/j.1365-3040.2007.01710.x ITEX Manual, updated 2011 Gus Shaver

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7496 gshaver@mbl.edu Lead PI Jim Laundre

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7496 arc_imr@mbl.edu Information Manager The goal of Arctic LTER project is to predict the future ecological characteristics of the site based upon our knowledge of the controls of ecosystem structure and function as exerted by physical setting and geologic factors, climatic factors, biotic factors, and the changes in fluxes of water and materials from land to water. To achieve this goal the Arctic LTER uses several approaches: Long-term monitoring and surveys of natural variation of ecosystem characteristics in space and time. Includes: climate, plant communities and productivity, thaw depth, stream flow, chemistry of streams and lakes, temperatures of streams and lakes, lake chlorophyll lake productivity, zooplankton abundance. Experimental manipulation of ecosystems for years and decades. Includes: tundra warming, shading, and fertilizing, grazer exclusions, fertilization of lakes and streams, addition and subtraction of predators. Synthesis of results and predictive modeling at ecosystem and watershed scales. Includes: stream N cycling, lake physics, bioenergetics of fish populations, water movement and transfer of DOC and nutrients from land to water, soil respiration, cycling and storage of C in tundra under different scenarios of future climates. This material is based upon work supported by the National Science Foundation under Grants #DEB-981022, 9211775, 8702328; #OPP-9911278, 9911681, 9732281, 9615411, 9615563, 9615942, 9615949, 9400722, 9415411, 9318529; #BSR 9019055, 8806635, 8507493. Any opinions, findings, conclusions, or recommendations expressed in the material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. The Arctic LTER research site (68°N and 149°W) is in the foothills region of the North Slope of Alaska and includes the entire Toolik Lake watershed and the adjacent watershed of the upper Kuparuk River, down to the confluence of these two watersheds. This area is typical of the northern foothills of the Brooks Range, with continuous permafrost, no trees, a complete snow cover for 7 to 9 months, winter ice cover on lakes, streams, and ocean, and cessation of river flow during the winter. Tussock tundra vegetation of sedges and grasses mixed with dwarf birch and low willows form the dominant vegetation type with areas of drier heath tundra on ridge tops and other well-drained sites as well as areas of river-bottom willow communities. -149.75 -149.0433 68.8 68.5 610 1360 meter 2012_GS_ITEX_PF_ShootHarvestData.csv Harvest data including the shoot leaf area index, position in the canopy, and shoot and plant tissue area, count and mass for each shoot harvested at three levels in the canopy from 19 1m x 1m plots dominated by S. pulchra and B. nana shrubs near LTER Shrub plots at Toolik Field Station, AK the summer of 2012. 2012_GS_ITEX_PF_ShootHarvestData.csv 1 \r\n column , " http://metacat.lternet.edu/das/dataAccessServlet?docid=knb-lter-arc.10142&urlTail=terrest/tracegas/data/2012_GS_ITEX_PF_ShootHarvestData.csv YEAR YEAR year of measurement yyyy DATE DATE date of measurement dd-mon-yy SITE SITE Toolik Toolik GROUP GROUP Measurement location in relation to Toolik Lake LTER Shrub plots; In vicinity of Block1 = Upland, IVO Block 2 = Outlet Measurement location in relation to Toolik Lake LTER Shrub plots; In vicinity of Block1 = Upland, IVO Block 2 = Outlet PLOT PLOT Individual plot identifier Individual plot identifier TREATMENT TREATMENT control or fertilized annually (with N and P) control or fertilized annually (with N and P) PLOT SIZE PLOT SIZE 1m x 1m point frame size 1m x 1m point frame size DOMINANT VEGETATION DOMINANT VEGETATION Dominant canopy vegetation Dominant canopy vegetation SPP SPP Shoot species sampled Shoot species sampled SHOOT ID SHOOT ID Identification for each shoot sampled (NOT unique); consists of canopy position (up/mid/low), species abbreviation, and a number Identification for each shoot sampled (NOT unique); consists of canopy position (up/mid/low), species abbreviation, and a number CANOPY POSN CANOPY POSN Relative position of shoot in the canopy Relative position of shoot in the canopy ROW# ROW# Row number on the point frame where pin-drop measurement were taken number real #N/A Missing or Not Measured PIN HOLE# PIN HOLE# Pin hole where pin-drop measurements were taken number real #N/A Missing or Not Measured DIST TO TOP OF SHOOT DIST TO TOP OF SHOOT Distance (cm) from the point frame to the highest tip of the shoot centimeter real #N/A Missing or Not Measured DIST TO BOTTOM OF SHOOT DIST TO BOTTOM OF SHOOT Distance (cm) from the point frame to the place point 5 inches from the tip centimeter real #N/A Missing or Not Measured DIST TO GROUND DIST TO GROUND Distance (cm) of the highest tip of shoot to the soil centimeter real #N/A Missing or Not Measured LAI (LAI-2000) LAI (LAI-2000) Leaf area index measured at shoot location (meter squared leaf per meter square ground) meterSquaredPerMeterSquared real #N/A Missing or Not Measured TOT SILHOUETTE AREA TOT SILHOUETTE AREA Area of whole shoot as viewed with shoot intact (square cm shoot) centimeterSquared real #N/A Missing or Not Measured SUM(GREEN LEAF, PETIOLE, STIPULE) AREA SUM(GREEN LEAF, PETIOLE, STIPULE) AREA Area of the shoot leaves, petioles, and stipules used to correct A/Ci and light curve data (square cm leaf+petiole+stipule) centimeterSquared real #N/A Missing or Not Measured GREEN LEAF AREA GREEN LEAF AREA Area of green leaves on shoot (square cm leaves) centimeterSquared real #N/A Missing or Not Measured PETIOLE AREA PETIOLE AREA Area of green petioles on shoot (square cm petiole) centimeterSquared real #N/A Missing or Not Measured STIPULE AREA STIPULE AREA Area of green stipules on shoot (square cm stipule) centimeterSquared real #N/A Missing or Not Measured OTHER GREEN ORGAN OTHER GREEN ORGAN Area of green organs on shoot (square cm organ) centimeterSquared real #N/A Missing or Not Measured BROWN STEM AREA BROWN STEM AREA Area of brown stem on shoot (square cm brown stemP centimeterSquared real #N/A Missing or Not Measured GREEN STEM AREA GREEN STEM AREA Area of green stem on shoot (square cm green stem) centimeterSquared real #N/A Missing or Not Measured TOTAL GREEN AREA TOTAL GREEN AREA Sum of green tissue area (leaf, petiole, stipule , organ, stem) on shoot centimeterSquared real #N/A Missing or Not Measured TOTAL SHOOT AREA (LEAVES, STEM, ORGAN) TOTAL SHOOT AREA (LEAVES, STEM, ORGAN) Sum of the area of all tissues (green and brown) on shoot (square cm shoot) centimeterSquared real #N/A Missing or Not Measured LEAF COUNT LEAF COUNT Number of leaves on shoot number real #N/A Missing or Not Measured PETIOLE COUNT PETIOLE COUNT Number of petioles on shoot number real #N/A Missing or Not Measured STIPULE COUNT STIPULE COUNT Number of stipules on shoot number real #N/A Missing or Not Measured GREEN ORGAN COUNT GREEN ORGAN COUNT Number of organs on shoot number real #N/A Missing or Not Measured TOT LEAF DRY MASS TOT LEAF DRY MASS Dry mass of all leaves (g leaf) gram real #N/A Missing or Not Measured TOT DRY PETIOLE MASS TOT DRY PETIOLE MASS Dry mass of all petioles (g petiole) gram real #N/A Missing or Not Measured TOT DRY STIPULE MASS TOT DRY STIPULE MASS Dry mass of all stipules (g stipule) gram real #N/A Missing or Not Measured AVG MASS PER LEAF AVG MASS PER LEAF Mass of leaves divided by leaf count (g per leaf) gramPerNumber real #N/A Missing or Not Measured AVG MASS PER PETIOLE AVG MASS PER PETIOLE Mass of petioles divided by petiole count (g per petiole) gramPerNumber real #N/A Missing or Not Measured AVG MASS PER STIPULE AVG MASS PER STIPULE Mass of stipules divided by stipule count (g per stipule) gramPerNumber real #N/A Missing or Not Measured AVG MASS PER ORGAN AVG MASS PER ORGAN Mass of organs divided by organ count (g per organ) gramPerNumber real #N/A Missing or Not Measured BROWN STEM DRY MASS BROWN STEM DRY MASS Dry mass of brown stem (g brown stem) gram real #N/A Missing or Not Measured GREEN STEM DRY MASS GREEN STEM DRY MASS Dry mass of green stem (g green stem) gram real #N/A Missing or Not Measured TOT SHOOT DRY MASS TOT SHOOT DRY MASS Dry mass of total shoot (g shoot) gram real #N/A Missing or Not Measured AVG AREA PER LEAF AVG AREA PER LEAF Total leaf area divided by leaf count (square meter per leaf) meterSquaredPerNumber real #N/A Missing or Not Measured LMA LMA Leaf mass per unit area (g leaf per square cm leaf) gramPerMeterSquared real #N/A Missing or Not Measured ORIGINAL LEAF MASS W/ PETIOLE ORIGINAL LEAF MASS W/ PETIOLE Dry mass of leaves with petioles prior to being separated and re-weighed (uncorrected g leaf) gram real #N/A Missing or Not Measured COMMENTS COMMENTS Notes on data quality and measurements Notes on data quality and measurements 116 2012_GS_ITEX_PF_ShootHarvestData.xlsx An excel file that has worksheets with the metadata and data. 2012_GS_ITEX_PF_ShootHarvestData.xlsx Microsoft Excel http://metacat.lternet.edu/das/dataAccessServlet?docid=knb-lter-arc.10142&urlTail=terrest/tracegas/xlsfiles/2012_GS_ITEX_PF_ShootHarvestData.xlsx Microsoft Excel file Leaf area in square meter per square meter of ground square centimeters Mass of leaves divided by number of leaves. Area of leaves divided by number of leaves grams per square meter