Table 3: The Growth Inhibition of the plb123 Mutant Upon AtPLA2 Expression is Principally the Result of Intracellular Accumulation of lysoPC. In Co-Cultivation Experiments, the plb123 Mutant was Transformed by Empty Vectors pYES2.1 and pESC with URA and LEU Selection Markers (Called plbEVEV), WT was Transformed by the Empty Vector (Called WTEV) or by PLA2 (Called WTPLA2) with the URA Selection Marker. The WTPLA2 was Then Co-Cultured with plbEVEV (WTEV Co-Cultured with plbEVEV was Used as the Control). Cells were Induced to Determine Whether WTPLA2 Could Release Enough lysoPC to Affect the Growth of plb123 Mutant Cells. In Order to Fully Reflect the PLA2 Effect, 10-Times the Number of WTPLA2 Cells and WTEV Cells were Grown Against plbEVEV Cells, which were Mixed Together for Co-Culture as Follows: After WTEV, WTPLA2 and plbEVEV were Cultured to OD600 3.0, WTEV and WTPLA2 Cells were Each Diluted to OD600 1.0 while plbEVEV Cells were Diluted to OD600 0.1 and Then the Preparations Co-Cultured. Then Either Before Induction or After Two Days Induction in the Presence of Galactose, Cultures were Diluted 100-fold and 10 µl was Spread on Plates with SC-URA-LEU Medium and Colonies Counted.
Co-Culture Strain Combination
Induction Time
# of Cells / mL Culture
WTEV + plbEVEV
0 hr
2.77E+05 ± 1.533E+04
WTPLA2 + plbEVEV
0 hr
2.83E+05 ± 5.774E+03
WTEV + plbEVEV
48 hr
1.40E+06 ± 1.447E+05
WTPLA2 + plbEVEV
48 hr
1.42E+06 ± 1.422E+05
Values are reported as # of colonies per mL of culture and are the average of three replicates ± SD.