This is a simple protocol for mating experiments in the Greenhouse at USU in July 2016.
We will have seperate cages for SD and LD treatments.
24 hours ahead of eclosion, we will move the pupae to the greenhouse. We will have cages set up the previous day.
Each cage will have 6 cups of flowers. Flowers will include alfalfa, lupin and some wildflowers.
Spray the cage with sugar water. Do not spray on the plastic side.
Hang tubes of sugar water in the cage, close to the opening side of the cage.
Place the pupa on a petridish in between the flower cups.
We don’t know yet if the pupae is male or female. So we will place around 6-8 pupae in one cage.
We will seal the zip side of the cage with duct tape. We will replace this tape regularly.
Collect eggs from each cage and bring them back to lab in a petridish and keep them under same light conditions as we kept them when we collected females from the wild.
Keep the diapause eggs in a petridish and leave them in the cabinet labelled “diapause eggs-July2016”.
Discard the new larvae.
Make sure there are no ants or any other critters in or around the cages.
Things to change regularly:
Flowers - change if you see any wilting
Sugar water : spray with sugar water and then regular water two times a day. Refill tubes with sugar water.
Duct Tape
Data to collect:
LD eggs:
Make note of dates on which eggs are collected.
If the eggs hatch into larvae : Count the number of eggs hatched on each day and record the date they hatched.
SD eggs:
Make note of dates on which eggs are collected.
If these eggs do not hatch in 3 days: we will consider them as “diapaused” eggs.
Count the number of eggs in diapause.
Count the number of eggs which hatch into larvae.
Record in spreadsheet:
Date of pupation for each larvae
Date of eclosion of each larvae
Seperate sheet with date and number of eggs collected for each treatment
LD: number of eggs hatched
SD: number of eggs in diapause and number of eggs hatched.