Controlled greenhouse phenotyping (Brassicaceae)

Knowledge graph centered on Controlled greenhouse phenotyping (Brassicaceae) with 59 nodes and 172 connections. Top connected: Ligusticum porteri, Atriplex canescens, Lepidoptera, Bacteria, Delphinium barbeyi.

Method synopsis

Standardized greenhouse cultivation of field-collected plant accessions under controlled temperature, light, and nutrition regimes for phenotype characterization.

Synthesized from method descriptions across 3 papers 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 — Ecological factors influence balancing selection on leaf chemical profiles of a wildflower (2021), Nature Ecology & Evolution. Implementations in other papers (listed below) may differ.

1. Generate near-isogenic lines (CFR-NILs)
   Wild accessions homozygous for production of methionine and branched-chain glucosinolates were identified and crossed. Multiple independent F3 families provide replicate homozygous MM and BB genotypes differing at BCMA1/3. Two homozygous F6 closest flanking recombinants (CFRs) were crossed together, yielding F1 and F2 CFR-NILs.
   Quantities: 6,860 CFR-NILs transplanted into field experimentsDuration: Multiple generations bred over several yearsConditions: Controlled greenhouse crosses
   Equipment: controlled greenhouse facilities
2. Establish field transplant arrays
   CFR-NILs were transplanted into 16 field environments across Colorado and Idaho. Plants were randomized within each site using a complete block design with at least eight CFR-NIL families to control for possible effects of unlinked loci outside the BCMA1/3 region. Arrays were arranged in three blocks, flush with the surrounding vegetation.
   Quantities: 16 environments, 6,860 total CFR-NILs transplantedDuration: Transplants monitored for 4-8 weeks in 2013 and 2015Conditions: Field conditions across elevational gradient in Rocky Mountains
   Equipment: field transplant materials
3. Measure fitness components in field
   We scored survival and herbivore damage weekly for the first two weeks of the growing season and then every other day. After eight weeks in field conditions, we censused for survival and herbivore damage. Herbivore damage was scored as percentage of leaf tissue damaged by insects.

Typical Equipment

• random number generator
• plant labels with unique IDs
• greenhouse environmental controls
• fertilization system

Output Measurements

• plant phenotype data
• chemical profiles

Papers Using This Protocol (3)

Species Studied With This Protocol