Spring.wmf (18300 bytes) Plant Physiology (Biology 327)  - Dr. Stephen G. Saupe;  College of St. Benedict/ St. John's University;  Biology Department; Collegeville, MN  56321; (320) 363 - 2782; (320) 363 - 3202, fax;    ssaupe@csbsju.edu

Protoplasts:  Pre-Lab Preparation

 I. Readings. To prepare for this lab you should read:

II. Additional Preparation. It would be helpful to review:

III. Questions: (Answer on a separate sheet; show your work)

  1. List the organelles you expect to see in a plant cell or protoplast with a light microscope.

  2. What determines plant cell shape? 

  3. What shape do you predict for the protoplasts? Why?

  4. How many grams of sucrose (MW = 342) is required to make 1.0 liter of a 0.5 M sucrose solution?

  5. How many grams of sucrose is required to make 1 liter of a 25% sucrose solution? What is the molarity of this solution?

  6. What is cyclosis and do you expect to observe it in the protoplasts?

  7. What types of contaminants do you expect to find in the initial protoplast suspension?

  8. Draw the chemical structures of sucrose, mannitol and glucose.

  9. Make a flow chart depicting, in general terms, a protocol that could be used to determine the concentration of nitrate in a leaf vacuole.

  10. The protoplast membrane is permeable to neutral red (True/False)

  11. The protoplast membrane is permeable to Evan's blue (True/False)

  12. The tonoplast is permeable to neutral red (True/False)

  13. Define: transvacuolar strand, cellulase, pectinase, hemicellulase.

  14. If the objects in Figure 1 in the hemocytometer write-up are chloroplasts, what is their concentration (chloroplasts � mL-1 )? Assume no dilution. SHOW YOUR WORK.
  15. Wagner (Plant Physiol. 64:88; 1979) reports that the average diameters of tulip leaf protoplasts and vacuoles are 80 �m and 75 �m, respectively.  (SHOW YOUR WORK)

16. Moskowitz and Hrazdina (Plant Physiol. 68:686; 1981) studied the vacuolar contents of cells from the skins of DeChaunac grapes. In their protocol, from 200 mature grape skins they obtained 1.5 X 107 vacuoles which was a yield of approximately 5%.  SHOW YOUR WORK.

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Last updated:  01/07/2009     � Copyright  by SG Saupe