Seed surface sterilization (Fabaceae)

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Method synopsis

Common garden experiment manipulating soil microbial communities through sterilization vs. inoculation to test effects on seed germination success and timing under controlled environmental conditions.

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 — Legume germination is delayed in dry soils and in sterile soils devoid of microbial mutualists: Species-specific implications for upward range expansions (2022), Ecology and Evolution. Implementations in other papers (listed below) may differ.

1. Soil sterilization and inoculation preparation
   Background soils were small-batch sterilized via double autoclaving within 12h at 121°C for 90min in 0.5-1.5L containers with drainage holes. Field-collected soils homogenized each year to standardize nutritional content and soil texture. For unsterilized treatments, added 15% field-collected soil inoculum to 85% sterile background soil and thoroughly mixed.
   Quantities: 0.5-1.5L containers, 121°C for 90min, 85% sterile background + 15% field inoculumDuration: 90min autoclaving, processed within 12hConditions: 121°C sterilization temperature
   Equipment: autoclave, Ray Leach Cone-tainer containers, Stuewe & Sons background soils (57% sand, 43% peat moss)
2. Experimental setup in growth chamber
   Sowed individual seeds directly into separate soil-filled pots at same depth, covered with ~10g treatment soil. Each treatment housed on separate tray to reduce microbe movement between pots. Placed in growth chamber (Conviron MTR30) at temperature and photoperiod reflecting early growing season at RMBL: day 21°C, 50% relative humidity, night 4.4°C, 20% relative humidity, 12:12h light:dark cycle.
   Quantities: ~10g soil cover per pot, 96 pots per tray, one seed per potDuration: 10 weeks monitoring periodConditions: Day: 21°C, 50% RH; Night: 4.4°C, 20% RH; 12:12h photoperiod
   Equipment: Conviron MTR30 growth chamber, 66ml pots with drainage holes

Typical Equipment

• autoclave
• Conviron MTR30 growth chamber
• Ray Leach Cone-tainer containers

Output Measurements

• germination success
• germination latency
• days to emergence

Papers Using This Protocol (1)

Species Studied With This Protocol

Places Where Used