NSF Project: Experimental
| Yeast Elicitation | UV Elicitation | Methyljasmonate Elicitation | Biological Tissue |
UV Irradiation Time Course
- Cultures were strained through a 3 inch strainer (American Houseware Mfg. Corp., Model #4332) by gravitation only (ie, no vacuum) such that the cells were still wet, and the cells were spread evenly on a Fisherbrand 150X15mm plate (Catalog #08-757-14) containing ~50ml SH agar using a Fisherbrand Disposable Cell Lifter spat (Catalog #08-773-1).
- After removing the lid, cultures were UV irradiated in a UV box for 5.5 minutes at 8000Joules/m2 , the lid was replaced and the plated cultures were stored upon a lighted shelf at 24C until time of harvest
- The cultures were 12P, or in the 12th passage of subculturing
- The time course was initiated on Wednesday, May 22, 2002, with the final 48 hour culture harvested on Friday, May 24, 2002
- The designated start time of the 0 time point and all subsequent time points is defined as the moment in which the UV treated plates came out of the UV box
- The 0 time point was initiated at 9:23AM; the final UV irradiated treatment was done by 11:29AM (2 hours and 6 minutes from treatment start to treatment finish)
- The methyljasmonate internal controls were initiated at 12:03, 12:05, and 12:06PM
- The yeast elicitor internal controls were initiated at 11:58AM, 12:00, and 12:00PM
- At time of harvest, the cells were gently scraped off the plate into the funnel and vaccuum applied; using the 50ml of 25% MS salts wash, the plate was rinsed to remove the remaining cells and the cells rinsed in the funnel as in the other time courses
- From this point, the cells were harvested and frozen as in the other time courses
- All time point harvests were relatively uneventful and as scheduled /planned
- All was well with this time course EXCEPT for the initial culture contamination and necessary adjustments made at the onset of the time course as described below.
For a detailed description of inoculum times and harvest times, see the attached spreadsheet.