cold stratification (Orobanchaceae)

Knowledge graph centered on cold stratification (Orobanchaceae) with 31 nodes and 109 connections. Top connected: Bombus, Ligusticum porteri, Bombus terrestris, Bombus appositus, Bombus impatiens.

Method synopsis

Cold stratification protocol for hemiparasite seeds followed by controlled germination under standardized temperature and photoperiod conditions. Includes seedling selection at specific developmental stage.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Parasitism modifies the direct effects of warming on a hemiparasite and its host (2019), PLOS ONE. Implementations in other papers (listed below) may differ.

1. Seed collection and storage
   Castilleja sulphurea mature fruits (capsules) were collected from 20 haphazardly chosen plants at 2900-3100 m near the Rocky Mountain Biological Lab (RMBL) in Gothic, Colorado, USA in September 2016. Bouteloua gracilis seeds collected in 2017 from Montezuma County, Colorado, USA at 1828 m were obtained from a local seed supplier (Western Native Seed, Coaldale, Colorado, USA). All seeds were stored in a 4°C refrigerator until processing.
   Quantities: 20 plants for C. sulphurea seed collection, seeds from 1828 m elevation site for B. gracilisDuration: Seeds collected in September 2016 for C. sulphurea, 2017 for B. gracilisConditions: Collection sites at 2900-3100 m and 1828 m elevation respectively, storage at 4°C
   Equipment: 4°C refrigerator
2. Seed stratification and germination setup
   C. sulphurea seeds were cold-stratified in spring 2018 by placing on moist filter paper in sealed Petri dishes in a 4°C refrigerator for ~120 days. Seeds were then sown in trays containing standard soil mixture (57% sand, 43% peat moss, and various minerals) and grown in a growth chamber at 21°C, 50% relative humidity and 4.4°C, 20% relative humidity on a 12:12 h light:dark cycle. Seeds were monitored daily for germination.
   Quantities: 120 day stratification period, 21°C day temperature, 4.4°C night temperature, 50% and 20% relative humidity, 12:12 hour photoperiodDuration: ~120 days for stratification, daily monitoring during germinationConditions: 4°C refrigerator for stratification, controlled growth chamber conditions
   moist filter paper, sealed Petri dishes, 4°C refrigerator, growth chamber (Conviron MTR30)

Typical Equipment

• 4°C refrigerator
• moist filter paper
• sealed Petri dishes
• growth chamber (Conviron MTR30)
• 1 L pots

Output Measurements

• germination rates
• seedling size
• leaf number

Papers Using This Protocol (1)

Species Studied With This Protocol