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(Data, Abstract, Methods, Variables)

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DATA SET IDENTIFIER: Interactions between plants and fungi and their roles in decay rates and CO2 release in five tropical leaf species

PROJECT TITLE: Decomposition Fungal - Plant Interactions (Factors influencing early-stage leaf litter decomposition in tropical forest with emphasis in the role of fungal – plant interactions )

PROJECT DESCRIPTION: Rates of decomposition depend on the particular interactions between producers and decomposer food webs.  This interactions are determined by the intrinsic characteristics of both plant and decomposers and are as well influenced by abiotic factors. Field studies were designed to determine the influences of environmental parameters such as climate and microsite variation on the decomposition rates of five tree species from The Luquillo Experimental Forest (LEF). This also allowed us to determine the influence of leaf quality on the decomposer community and whether leaf physical, structural or phylogenetic relationships could be used as predictors of decomposition rates. Microcosms were used to separated the contribution of interactions between dominant plants and fungal species in decomposition rates. The rates of decomposition were determined by mass loss in both the field and microcosms experiments.  In the microcosms systems, CO₂ evolution by the decomposition of each of the five leaf litter substrates, by fungal species isolated from that particular leaf or from any of the other litter types, was obtained by using sodium hydroxide traps. There were significant differences in decomposition rates among leaf litter types both in field and microcosms experiments.  There was also a significant effect of dry and wet periods in decomposition rates.  Leaf litter decomposed faster under their  tree sources than in a common plot.  Among the leaf quality parameters, the lignin and nitrogen to lignin ratios were the best predictors of decomposition rates. Non polar elements, water soluble (simple sugars), P and Ca were positively correlated with percent mass loss (PML) while C was negatively correlated with PML. In the microcosms experiments we did not found specificity between fungi and their substrate (where the fungus was isolated), neither to substrates that were chemically, physically or structurally related to the plant were the fungi was originally isolated. Nevertheless, there were differential responses of particular fungi to plant substrates as well as influences of plant species on the fungal decomposers performance.  The differential contributions of leaf species to carbon budgets (mass remaining and CO₂ release) may be important in determining management practices for forest and agricultural systems. We found that species such as Manilkara bidentata  may retain carbon in the ecosystem while other species such as Sapium laurocerasus  decomposed rapidly and therefore, released nutrients quickly but with greater carbon losses. The absence of tight links between plants and fungal decomposer may indicate adaptation of fungi to changes in resource availability in a disturbed forest. Alternatively, this might indicate that the presence of a fungi in a particular substrate does not depend on the substrate's chemical o physical composition but on the presence of other members of the detrivoral community.  Although the presence of generalist basidiomycetes made an important contribution to the decomposition of leaf species, the diversity of the decomposer and detrivoral community might be important in maintaining nutrient balances in the ecosystems since random encounters of plants, decomposers and detrivores may determine the residence time of a substrate on the forest floor.

LTER CORE AREAS: (Annotate all that apply)

LEF LTER 1 RESEARCH TOPIC: (Annotate all that apply)

We define a data file as a component of a data set. A data set can have only one data file or more. Basically, different data files have different data structures or format.
DATA SET FILES (SUBSETS):

RESEARCH LOCATION: Recently fallen leaves of five tree species were collected at three sites in a non-seasonal, late secondary, subtropical wet forest.  The sites were El Verde, EV (3500 mm ppt y-1; 350 masl), Sabana, Rte. 988 (3000-4000 mm ppt y-1; 250 masl) and Bisley (3000 mm ppt y-1; 150-250 masl); all sites are in the Caribbean National Forest, Puerto Rico.

INVESTIGATORS:

OTHER RESEARCHERS	E-MAIL address
D. Jean Lodge	djlodge@caribe.net

SOURCE OF FUNDING (SPONSOR): NSF - LEF LTER

DATA SET ABSTRACT: A microcosm experiment was used to test for the effects of interactions between particular plant and fungal decomposer species on rates of leaf decomposition. Each microcosm contained one species of leaf that was sterilized with gamma irradiation and then inoculated with a single fungus. Five plant species and ten fungal species (two dominants from each of the litter types) were used in all possible combinations. Plant species were selected for pair-wise comparisons based on phylogenetic relationships and litter quality characteristics. Decomposition was measured by both mass loss and CO₂ release. Differences in weight loss and CO₂ evolution were highly significant for plants, fungal species, and their interactions. Mass loss was positively correlated with CO₂ evolution. Contrary to our hypotheses, however, microfungal dominants did not decompose their source leaves faster than microfungal dominants from other leaf species, nor were responses to other types of specificity detected. Matching of fungi to leaf substrates by their source, by phylogenetic relationships, or by chemical, physical and structural characteristics was not associated with consistent increases in decomposition. Although previously documented differences in microfungal species composition and dominance among decomposing leaves of different trees were confirmed in this study, such differences apparently do not directly affect the rates of ecosystem processes. The presence in a few of the microcosms of a generalist basidiomycete that had ligninolytic enzymes, Melanotus eccentricus, significantly accelerated the rate of decomposition. Non-specific basidiomycetes may therefore have a stronger effect on early stages of leaf litter decomposition than host-selective microfungi.

DATA SET METHODS:
Methods
Plant species selection
Plant species were selected for pair-wise comparisons based on phylogenetic relatedness and structural characteristics (e.g. lignin and thickness) and chemical composition, (nitrogen and phosphorus concentrations). Croton poecilanthus Urban and Sapium laurocerasus Desf. both belong to the Euphorbiaceae, but they differ in physical and chemical characteristics. Croton poecilanthus and Manilkara bidentata (A.DC.) Chev. leaves have similar physical and chemical traits including the presence of latex, though the latter belongs to a different family (Sapotaceae).  Inga vera Willd. and I. fagifolia (L.) Willd. were selected to represent two closely related species in the Leguminoseae with high lignin and nitrogen concentrations, but differing slightly in concentrations of secondary plant compounds.

Leaf  collection
Recently fallen leaves of five tree species were collected at three sites in a non-seasonal, late secondary, subtropical wet forest.  The sites were El Verde, EV (3500 mm ppt y-1; 350 masl), Sabana, Rte. 988 (3000-4000 mm ppt y-1; 250 masl) and Bisley (3000 mm ppt y-1; 150-250 masl); all sites are in the Caribbean National Forest, Puerto Rico. Leaves of M. bidentata and C. poecilanthus were all collected from EV, I. fagifolia and I. vera leaves were collected at EV and Sabana, and S. poecilanthus leaves were collected at EV and Bisley.  Leaves of each species were taken from at least five trees that were growing at least 20 m apart at each site.

Determination of dominant early decomposer fungi by the particle filtration method
Decomposed leaves were collected and air dried for three hours before microfungal species were isolated using the particle filtration method as described by Bills and Polishook (1994, 1996) and Polishook et al. (1996), and modified by Bills (2000). The decomposed leaves were placed in a sterile blender and pulverized at high speed for one minute. Pulverized samples were washed with a stream of sterile distilled water (10 min) through a 2 mm brass pre-screen, and two sterilized polypropylene mesh filters (210 and 105 µm) to remove spores. The particles trapped on the 105 µm mesh filter were placed in a 50 ml sterile polystyrene centrifuge tube and 50 ml of sterile distilled water was added to re-suspend them. After letting the particles settle, the water was decanted to remove more spores and any remaining water was extracted with a sterile pipette. Particles were washed again with a second volume of water to obtain a 20:1 dilution; this procedure was repeated again twice. Washed particles were diluted to 1:100, 1:5000 and 1:2,000,000 using a serial dilution method. From each serial dilution, 0.1 ml was plated and evenly distributed on the agar surface in 90 mm petri dishes using a flamed, bent-glass rod (10 plates each of Malt-Cyclosporin Agar and Bandoni’s Medium, as described below). This procedure was carried out in a sterile hood. The plates were incubated at room temperature (ca. 22 C) with a 12 h photoperiod.

Two types of culture media were used for the initial dilution plates, Malt-Cycloporin Agar (Malt Yeast Agar, as described below, with 10 mg of Cyclosporin A added when the medium was cool; Polishook et al. 1996) and Bandoni's Medium (4g of L-sorbose, 0.5 g yeast extract and 20 agar L-1 distilled water). Fifty mg L-1  of Chlortetracycline and Streptomycin Sulfate were also added to the Malt-Cyclosporin Agar and Bandoni's Medium when the agar was cool to prevent bacterial growth. Fungi growing from the particles were transferred to 90 mm petri dishes with Malt Yeast Agar (MYA: 10 g of malt, 2g yeast extract and 20 g agar per L distilled water). Oatmeal (OMA), Corn Meal (CMA), Malt (MA) and Potato Dextrose Agar (PDA) were made according to Rossman et al. (1998), and were used for separation and identification of strains.

All fungi growing from particles were isolated. Growing fungi were transferred to duplicate slants of MYA and morphologically similar species (morphospecies) were sorted after one month of growth. Sub-samples of similar species and species that did not sporulate were transferred to slants of OMA, CMA, PDA and MEA to verify that the morphospecies were correctly classified.  Autoclaved banana leaves, and in some cases, autoclaved leaves of the species from which the fungi were originally isolated, were added to PDA and MEA media to promote fungal sporulation. These cultures were allowed to grow for another month and sorted. After two months, morphologically similar species were sorted again and their frequencies were recorded.

Determination of maximum likehood of fungal diversity to adjust methods
A trial experiment to isolate fungal species from S. laurocerasus leaves was set up in order to determine the dilution that yielded the maximum diversity of fungal species and would therefore allow the best possible representation of the fungal community. Sapium laurocerasus was chosen for the trial because it is known to have high litter quality and be easily decomposable (Harmon 2000), and can therefore be expected to support an initially high diversity of fungi.
Freshly fallen senescent S. laurocerasus leaves were collected at El Verde, surface dried with paper towel, placed in litter bags, returned to the same site, and decomposed for five weeks (May 13 to June 16, 2000).  The particle filtration method described above in section 2.3. was used except that only 3.75 g of air dry leaves were available instead of the recommended five grams. The maximum number of strains isolated using each medium was considered the maximum expected number of species that we could potentially isolate. Previous research on two litter species, Manilkara bidentata and Guarea guidonea (L.) also provide information on microfungal diversity in leaf litter species at El Verde, but the diversity was higher in that study because litter at various stages of decomposition was used (Polishook et al. 1996).

Determination of the dominant fungi in each leaf species
In order to determine the ten vegetatively dominant fungal species among the early stage decomposers in each leaf litter species, a five-week decomposition experiment was established underneath five representative trees per plant species. Five grams of senescent freshly fallen leaves of each species were placed in separate 20 cm2 mesh bags. Five bags per species were placed under each of the five tree replicates of the same species (25 bags per tree species). Leaf litterbags were collected after five weeks of decomposition to isolate early stage fungal decomposers. One litterbag per tree replicate was randomly selected for fungal isolation among the original five and these were pooled for isolation of fungi. Isolations were made from a 5 g subsample from the pooled litter sample. The random selection of one bag from each of the five tree locations allowed us to adjust for heterogeneity within tree species.
Fungi were isolated using the particle filtration method and sorted into morphospecies as described above in section 2.3, except that only five of the ten plates were randomly selected from the 1:100 dilution as the source of the culture isolates. A total of 182-294 cultures were obtained from each of the leaf species (Table 1). Classification of the morphotypes was reviewed at two and four months, and the total number of isolates per morphospecies was recorded. The most common species were identified to genus when possible.

Selection of fungal isolates for the microcosm experiment
For each of the leaf species, the fungal morphotypes were arranged by their rank abundance based on frequencies of occurrence, and selections were made from the five most frequent morphotypes. In general, two dominant fungal species were selected from each leaf species that did not occur among the dominants in the other leaf litter species. While the frequency data from this study (Table 1) confirm previous findings of differences in fungal community composition and dominance among litter species (Holler and Cowley 1970, Cornejo et al. 1994, Polishook et al. 1996), there was some overlap in species composition among dominant fungi from the five leaf types. Fusarium solani the most frequent fungus from S. laurocerasus was also frequent in I. fagifolia. Fusarium was selected to represent S. laurocerasus since two other dominants (Trichoderma sp. and Penicillium) were eliminated based on their ability to produce airborne spores whichmakes them dangerous to health and difficult to control in terms of cross-contamination (Table 1). Similarly, Volutella sp. was the second most frequent isolate from C. poecilanthus leaves, but it also occurred among the top five dominants in S. lauracerasus (pre-trial), and also among the five most frequent species in M. bidentata (Table 1). The most frequent isolate from M. bidentata in a previous study (Polishook et al. 1996) as well as in this study, Pestalotia sp., was used although it was also among the five most frequent species in C. poecilanthus leaves. Pestalotia species are common but difficult to identify, and the species from M. bidentata and C. poecilanthus may or may not be the same. Plant species were assigned a code corresponding to the first letter of genus and species. Fungal species were assigned the plant code from their source leaf followed by the number 1 or 2 according to their relative ranks. For example, the two isolates from M. bidentata were coded as MB1 and MB2. The selected fungal isolates and the other eight most frequent isolates from each leaf litter species appear in Table 1.

Microcosm design
Translucent plastic containers (19 cm x 12 cm and 8.5 cm deep; 1892 ml volume, Glassware™) were modified for use as microcosms. A ventilation tube made from a 7 cm length of plastic pipette (1 ml) allowed gas exchange. The ventilation tube was sealed to the microcosm with hot glue, and one cm extended inside the container. The ends of the ventilation tubes were covered with two layers of parafin film (Parafilm) to prevent entrance of fungal spores and mites. A wire mesh platform (1 cm mesh) was inserted to form a shelf 4 cm from the bottom of each microcosm. An autoclaved piece of cloth was placed over the wire platform to prevent litter from falling through the mesh. A gap in the wire mesh in one corner allowed insertion of vials containing a solution to trap CO2 . The microcosms were placed in a laminar flow hood and misted with 70 ml of sterile deionized water once per week.

Leaf sterilization
Senescent freshly fallen leaves were collected and sterilized with gamma irradiation (3640 rads of Cs137). Culturing from those leaves showed that viable propagules had survived the first sterilization. A second, stronger irradiation was therefore performed a month later (500 rads/min Cs137 exposure time 8.18 min total radiation 4090 rads). No colonies grew from the subsamples that were cultured after the second irradiation.

Leaf inoculation
Five grams of leaf litter of a single plant species were placed in each microcosm. Leaves in each microcosm were inoculated with only one fungal species. The five leaf species were inoculated with two dominant decomposer fungi from each of the five leaf species in all possible combinations. There were three replicates for each of the 50 treatment combinations for a total of 150 microcosms. Sterile leaves were placed in the microcosms, moistened with sterile distilled water, and inoculated with 10 agar plugs (5 mm dia) from clean cultures. In addition, 1 ml of liquid medium containing the fungus was added to the leaves to ensure rapid fungal colonization.
The numbers for the 150 microcosms were assigned randomly to randomize the placement of treatment combinations during incubation. When measurements of CO2 were made, however, the microcosms were divided into two groups to allow better management and to insure that treatment combinations that were critical for testing the original hypotheses were measured on the same day. Group I contained S. laurocerasus, C. poecilanthus and M. bidentata for phylogenetic relatedness versus litter quality contrasts. Group II was inoculated on the following day, and was comprised of the two Inga species.

Re-isolation of fungi
Subsamples of litter from each microcosm were cultured on MEA to determine the fungi present at the end of the experiment. Some microcosms containing leaves of Croton poecilanthus were found to have agaric basidiomycetes with ligninolytic enzymes in addition to the microfungal isolate in the treatment. One microcosm had a Marasmius sp., while the others with basidiomycetes had Melanotus eccentricus (Murrill) Sing. This apparently resulted from incomplete gamma sterilization of Croton leaves. Comparison of weight loss in replicates with and without basidiomycetes indicated significant differences in weight loss (t-tests). Of the 150  microcosms, 16 were found to have microfungal contaminants at the end of the experiment. (Appendix Table 1). These were probably contaminated late in the experiment, and values for weight loss and CO2 evolution did not differ between contaminated and uncontaminated replicates (t-tests). Data from microcosms with microfungal contaminants were therefore treated according to the fungi in the original treatment, while those with delignifying basidiomycetes were treated separately in statistical analyses.

CO2 sampling
Traps that contained 15 ml 1 M NaOH were placed in the microcosms, which were then closed tightly for 24 h to measure the rates of CO2  evolution. Samples were collected during wks 1, 2, 3,4, 8 and 17. Group II was processed one day after Group I on each sample date so that the number of days of incubation was the same for both groups. After each collection, a 5 ml subsample from the NaOH-CO2 trap was combined with 2.5 ml of  BaCl2 (1M) and 2 drops of Phenolphthalein. The subsample was then titrated with HCl (1N). The volume of HCl consumed was used to calculate the amount of CO2  in the trap solution. Mean daily CO2 evolution on each of the four sample dates was calculated from the replicates for each plant-fungus combination. Mean daily CO2 evolution g-1 air dried litter across sample dates was also calculated.

REFERENCES:
Bills, G. F. 2000. Introduction to techniques for collection of soil and litter fungi. British Mycol. Soc. Millenium Symposium on Tropical Mycology, April 2000. John Moore University, Liverpool, UK. 1-24.

Mikola, J., and  Setälä, H. 1998. Productivity and trophic level biomasses in a microbial-based soil food web. Ecology 79, 153-164.

Polishook J. D., Bills, G. F. and Lodge, D. J. 1996.  Microfungi from decaying leaves of two rain forest trees in Puerto Rico. Journal of Industrial Microbiology 17, 284-294.

Taylor B. R and Parkinson, D. 1988a. Respiration and mass loss rates of aspen and pine leaf litter decomposing in laboratory microcosms. Canadian Journal of Botany 66, 1948-1959.

Taylor B. R and Parkinson, D. 1988b. Annual differences in quality of leaf litter of aspen (Populus tremuloides) affecting rates of decomposition. Canadian Journal of Botany 66, 1940-1947.

CROSS-REFERENCES (other data sets related to this one): LTERDBAS#124: Factors influencing decomposition of leaves for five plant species at El Verde

SAMPLE LOCATION: ITES Facundo Bueso 104, Rio Piedras Campus, University of Puerto Rico

STORAGE SITES (of data files): LUQ LTER Data Manager's Cabinet DM333-00, Drawer #4; USDA Forest Service Sabana Luquillo

INVESTIGATOR'S ASSIGNED KEYWORDS: Decomposition, leaf litter, microfungi, basidiomycetes, CO₂, tropical forest, microcosms

LEF LTER OFFICIAL KEYWORDS (See table): EL VERDE, TABONUCO, BIOGEOCHEMICAL CYCLES, CARBON, NITROGEN, DECOMPOSITION, LITTER FALL, SECOND FOREST,  ASCOMYCETIES, BASIDIOMYCETES, THESIS

PUBLICATIONS:
Santana. M.E., D.J. Lodge and Lebow. P. 2005. Relationship of host recurrence in fungi to rates of tropical leaf decomposition. Pedobiologia 49 (2005) 549-564.

DISSEMINATION: RESTRICTED (until the two year period)

REASONS TO RESTRICT DATA IN THIS DATA SET BEYOND ITS TWO YEAR POLICY PERIOD

*:*WILL HAVE TO BE APPROVED BY LTER PRINCIPAL INVESTIGATORS: N. BROKAW, J. ZIMMERMAN, A. LUGO , W. MCDOWELL, W. SILVER

SITES DESCRIPTIONS:

Geographical positional system (GPS) Coordinates for each location:

VARIABLES (ATTRIBUTES):

FILE NAME OR #ABOVE (all in which the variable appears)	1,2	1,2	1,2	1,2	1,2
ABBREVIATION (as it appears on the data file)	Plantspecies	Microcosm	Combination	Plantcode	Fungalspecies
NAME OF VARIABLE	Species name of plant	Microcosm code number	Combination code	Code of plant  species	Species name of fungus
DEFINITION OF VARIABLE	Species name of the leaves used in the experiments	Random number assigned the combination of plant and fungal species	Plant and fungal species combination code	Code assigned to each species	Fungal species inoculated on five grams of leaf litter of a single plant placed in each microcosm
UNIT
PRECISION
RANGE OR LIST OF VALUES	Croton poecilanthus, Inga fagifolia, Inga vera, Manilkara bidentata, Sapium laurocerasus	{1,…,150}	CP-IF1 = Croton poecilanthus,Aureobasidium cf. pullulans, CP-IF2 = Croton, poecilanthus,Cladosporium sp., CP-MB2 = Croton poecilanthus,Colletotrichum sp. 2, CP-CP2 = Croton poecilanthus,Diplodia sp., CP-SL1 = Croton poecilanthus,Fusarium solani, CP-SL2 = Croton poecilanthus,Mucor sp., CP-IV1 = Croton poecilanthus,Nectria sp. 2, CP-MB1 = Croton poecilanthus,Pestalotia sp., CP-IV2 = Croton poecilanthus,Phoma sp., CP-CP1 = Croton poecilanthus,Vollutela cf. concentrica, CP-CP2 = Croton poecilanthus,Vollutela cf. concentrica, IF-IF1 = Inga fagifolia,Aureobasidium cf. pullulans, IF-IF2 = Inga fagifolia,Cladosporium sp., IF-MB2 = Inga fagifolia,Colletotrichum sp. 2, IF-CP1 = Inga fagifolia,Diplodia sp., IF-SL1 = Inga fagifolia,Fusarium solani, IF-SL2 = Inga fagifolia,Mucor sp., IF-IV1 = Inga fagifolia,Nectria sp. 2, IF-MB1 = Inga fagifolia,Pestalotia sp., IF-IV2 = Inga fagifolia,Phoma sp., IF-CP2 = Inga fagifolia,Vollutela cf. concentrica, IV-IF1 = Inga vera,Aureobasidium cf. pullulans, IV-IF2 = Inga vera,Cladosporium sp., IV-MB2 = Inga vera,Colletotrichum sp. 2, IV-CP1 = Inga vera,Diplodia sp., IV-SL1 = Inga vera,Fusarium solani, IV-SL2 = Inga vera,Mucor sp., IV-IV1 = Inga vera,Nectria sp. 2, IV-MB1 = Inga vera,Pestalotia sp., IV-IV2 = Inga vera,Phoma sp., IV-CP2 = Inga vera,Vollutela cf. concentrica, MB-IF1 = Manilkara bidentata,Aureobasidium cf. pullulans, MB-IF2 = Manilkara bidentata,Cladosporium sp., MB-MB2 = Manilkara bidentata,Colletotrichum sp. 2, MB-CP1 = Manilkara bidentata,Diplodia sp., MB-SL1 = Manilkara bidentata,Fusarium solani, MB-SL2 = Manilkara bidentata,Mucor sp., MB-IV1 = Manilkara bidentata,Nectria sp. 2, MB-MB1 = Manilkara bidentata,Pestalotia sp., MB-IV2 = Manilkara bidentata,Phoma sp., MB-CP2 = Manilkara bidentata,Vollutela cf. concentrica, SL-IF1 = Sapium laurocerasus,Aureobasidium cf. pullulans, SL-IFI = Sapium laurocerasus,Aureobasidium cf. pullulans, SL-IF2 = Sapium laurocerasus,Cladosporium sp., SL-MB2 = Sapium laurocerasus,Colletotrichum sp. 2, SL-CP1= Sapium laurocerasus,Diplodia sp., SL-SL1 = Sapium laurocerasus,Fusarium solani, SL-SL2 = Sapium laurocerasus,Mucor sp., SL-IV1 = Sapium laurocerasus,Nectria sp. 2, SL-MB1 = Sapium laurocerasus,Pestalotia sp., SL-IV2 = Sapium laurocerasus,Phoma sp., SL-CP2 = Sapium laurocerasus,Vollutela cf. concentrica	CP = Croton poecilanthus, IF = Inga fagifolia, IV = Inga vera, MB = Manilkara bidentata, SL = Sapium laurocerasus	Aureobasidium cf. pullulans, Cladosporium sp., Colletotrichum sp. 2, Diplodia sp., Fusarium solani, Mucor sp., Nectria sp. 2, Pestalotia sp., Phoma sp., Vollutela cf. concentrica
DATA TYPE	alphabetic	integer	alphanumeric	alphabetic	alphanumeric
MISSING DATA CODES	none	none	none	none	none

VARIABLES (ATTRIBUTES):

VARIABLES (ATTRIBUTES):

FILE NAME OR #ABOVE (all in which the variable appears)	2	2	2	2	2
ABBREVIATION (as it appears on the data file)	Funguscode	Plantnumber	Fungusinoculumcode	Basexclusion	Basdummy
NAME OF VARIABLE	Code of fungal species	Number code of species	Fungus inoculum code	Fungi inoculum and basidiomycete combinations	Basidiomycete dummy variable
DEFINITION OF VARIABLE	Code assigned to the fungal species inoculated on five grams of leaf litter of a single plant placed in each microcosm	Code assigned to each species	Number code assigned according to the plant source from which the fungus was isolated	Fungal inocula and basidiomycete combinations	Dummy variable representing present or absense of the basidiomycete
UNIT
PRECISION
RANGE OR LIST OF VALUES	IF1 = Aureobasidium cf. pullulans, IF2 = Cladosporium sp., MB2 = Colletotrichum sp. 2, CP1 = Diplodia sp., SL1 = Fusarium solani, SL2 = Mucor sp., IV1 = Nectria sp. 2, MB1 = Pestalotia sp., IV2 = Phoma sp., CP2 = Vollutela cf. concentrica	4 = Croton poecilanthus, 2 = Inga fagifolia, 1 = Inga vera, 3 = Manilkara bidentata, 5 = Sapium laurocerasus	IV1 = 1, IV2 = 2, IF1 = 3, IF2 = 4, MB1 = 5, MB2 = 6, CP1 = 7, CP2 = 8, SL1 = 9, SL2 = 10	IV1 = 1, IV2 = 2, IF1 = 3, IF2 = 4, MB1 = 5, MB2 = 6, CP1 = 7, CP2 = 8, SL1 = 9, SL2 = 10, B -IV1 = 11, B-F1 = 12, B-MB2 = 13, B-CP1 = 14 (B = Basidiomycete)	0 = absent, 1 = present
DATA TYPE	alphanumeric	integer	integer	alphanumeric	decimal
MISSING DATA CODES	none	none	none	none	none

VARIABLES (ATTRIBUTES):

FILE NAME OR #ABOVE (all in which the variable appears)	2	2	2	2
ABBREVIATION (as it appears on the data file)	Time(weeks)	CO2time17wk	CO2mg/g-d	CO2mg/5g-dmicrocosm
NAME OF VARIABLE	Time (in weeks) allowed for decomposition	CO2 concentration measured at collection time	Mean CO2 milligram per day	CO2 milligrams produced
DEFINITION OF VARIABLE	Number of weeks the sample were allowed to decompose in the field before collection	CO2 concentration measured at time of collection after the time (17 weeks) they were allowed to decompose	CO2 milligrams per gram of leaf litter divided by all day collections	CO2 milligrams produced by 5 grams of leaves and inoculum
UNIT		milligrams	milligrams	milligrams
PRECISION		.1	.1	.1
RANGE OR LIST OF VALUES	17.00
DATA TYPE	integer		decimal
MISSING DATA CODES	none		none

COMPUTATIONAL METHODS:

Variable Name	Formula
Percent mass remaining	[Initial Mass(g)/ Final Mass(g)]/100
Percent leaf mass lost	100 - Percent mass remaining

FOR DATA MANAGER USE

DATE OF LAST REVIEW: November 4, 2008
DATE OF LAST ENTRY : November 17, 2000
STAGE OF DATA SET MANAGEMENT (dates) :
RECEIVED ENTERED: September 27, 2002
FILED ON-LINE April 1, 2003 (metadata only); April 29, 2008 (with data)
REVIEWED BY RESEARCHER
FILING MEDIA :
NAME OF DOCUMENTATION FILE : lterdb125.doc.htm
NAME OF ON - LINE CATALOG : LTERDBAS
RECORD # :   125
DOCUMENT TYPE : data set (magnetic media)
PRIORITY TO BE ENTERED :  N/A

Rev. date of this form: 8 June 2001 / 20 May 2003