Hierarchical leaf trait sampling in aspen clones (Salicaceae)

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Method synopsis

Hierarchical sampling design with nested random effects (site, plot, tree, branch) to assess intraspecific trait variation while accounting for clonal structure. Multiple leaves collected per branch to capture within-tree variation.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure as described in the canonical source

Steps below were extracted from the paper that introduces this protocol — Relaxation of the leaf economics spectrum within and across quaking aspen Populus tremuloides genotypes (2025), Oikos. Implementations in other papers (listed below) may differ.

1. Establish study sites and plots
   Selected four sites along an elevational gradient in Gunnison National Forest near RMBL: Deer Creek (3000-3250 m), RMBL (2900-3150 m), Avery (3100-3300 m), and Bellview (3270-3635 m). At each site, established five plots with continuous aspen stands along south- to southwest-facing slopes with continuous aspen stands along an altitudinal gradient.
   Quantities: 4 sites, 5 plots per site, distances between sites ranging from 1.5 to 5 kmDuration: Not specifiedConditions: South- to southwest-facing slopes, elevational range 2900-3635 m a.s.l.
   Equipment: GPS for site location
2. Select and mark trees within plots
   At each plot, selected ten trees per plot, two branches per tree, and four leaves per branch for leaf trait measurements. Central tree was selected for each plot and used for stratified random selection of surrounding 10 study trees using point polar method. Plots were designed with a hierarchical sampling strategy to assess trait-trait relationships within and across aspen genotypes.
   Quantities: 10 trees per plot, 2 branches per tree, 4 leaves per branch, total 172 trees yielding 344 branchesDuration: Not specifiedConditions: During growing season
   Equipment: Point polar method equipment
3. Collect leaf samples for trait measurements
   Eight healthy adult leaves from two branches per tree (four leaves per branch) were selected for SLA and LDMC measurements. Leaves were collected and dried at 60°C for 48 h and then weighed again. For LNC analysis, 10 g of dried soil was used for GWC analysis in cases where soil sample was less than 10 g.

Typical Equipment

• portable photosynthesis system
• drying oven
• analytical balance
• pH meter

Output Measurements

• SLA
• LDMC
• LNC
• leaf lifespan
• photosynthetic capacity
• dark respiration

Papers Using This Protocol (1)

Species Studied With This Protocol