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Cell Biology JoVE (Open Access) Citable · DOI

Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses

DOI: 10.3791/50527-v
What you'll learn
  • ✓ Establish controlled growth conditions for Setaria viridis flowering
  • ✓ Perform panicle emasculation using hot water treatment
  • ✓ Execute cross-pollination and identify successfully outcrossed progeny
Protocol

We have developed a methodology for performing crosses in Setaria viridis (S. viridis). The method involves pruning the panicle prior to a hot water treatment to kill viable pollen. Crosses are performed following a well-controlled growth regime and typically result in the recovery of 1 to 7 cross-pollinated seed/s per panicle.

Difficulty
intermediate
Total time
~2–3 weeks (plant growth + crossing + seed maturation)
Model organism
Setaria viridis

Steps

1
Grow Setaria viridis under controlled conditions

Establish standardized growth regime to produce flowering plants with mature panicles ready for crossing.

▶ 01:10
2
Trim and label panicle before emasculation

Prune the panicle to remove excess florets and apply identification labels for tracking crosses.

▶ 02:11
3
Emasculate panicle with warm water dipping

Immerse prepared panicle in hot water to kill viable pollen and prevent self-fertilization.

▶ 03:49
4
Perform cross-pollination and harvest seeds

Apply pollen from donor plant to emasculated panicle and collect resulting cross-pollinated seeds.

▶ 04:47
5
Identify successfully outcrossed progeny

Analyze seed and seedling characteristics to confirm successful cross-pollination events.

▶ 07:32
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