Turnip (Brassica rapa L.) root tissue was exposed to freeze-thaw stresses of −7, −9, -−11 and −19 °C. The post-thawed tissues were either subjected to leaching in deionized water for 2 h or left at 100% humidity. Tissue survival was then assayed by vital staining using modified 2,3,5-triphenyltetrazolium chloride (TTC) staining in 0.2 M phosphate buffer. Tissue survival was significantly lower for leached samples than for non-leached samples. It is concluded that freeze-thaw injury in plant tissues is enhanced by post-thaw leaching in water. The 0.05 M phosphate buffer commonly used for TTC staining also damaged freeze-thaw injured tissue. Key words:Brassica rapa L., 2,3,5-triphenyltetrazolium chloride (TTC), freeze-thaw injury, leaching
This laboratory has been using a derivatized bead assay to study cell surface properties in many experimental systems. Here we test its efficacy in the yeast/concanavalin A (Con A) binding model to determine if immobilized Con A is inhibited by the same saccharides that inhibit free Con A binding. This system offers an excellent test case for the efficacy and validity of the derivatized bead assay. Con A binds to yeast because yeast are rich in cell surface mannose residues, a Con A preferential binding sugar. Here we examine the effects of 30 different sugars (0.05 M) on the binding of yeast to agarose beads derivatized with Con A, in a total of 3998 trials, with an average of over 133 replicates for each sugar. The most inhibitory sugars included D(+) melezitose, D(+) trehalose and maltotriose, the same sugars that effectively inhibit binding of free Con A. The results suggest that the bead assay is an effective approach to study the binding properties of cells, and most important, it is so rapid that hundreds of trials can be done in the time it would take to do one trial using conventional assays (supported by NIH NIGMS SCORE, RISE, MARC, the ITQ program, and the Joseph Drown Foundation).
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