Figure 4.

Competitive PCR Analysis of CRY mRNA Accumulation during Light-Dependent Spore Germination.

The amounts of each CRY transcript were determined by using competitive PCR. Spores imbibed in the dark were transferred to either red light or blue light. The amounts of CRY transcripts were measured at various times after transfer. This experiment was repeated twice in its entirety with very similar results in each case.

(A) Representative gel images of competitive PCR for CRY4 and CRY5. The PCR products derived from cDNA (closed arrowheads) and competitor (open arrowheads) are indicated on the gel image at each 1-hr red light irradiation sample after 4 days of incubation in the dark. The copy numbers of the competitors (from 10¹ to 10⁸ copies) in each reaction tube are shown at the top of the sequential gel images. DS, dry spores; 1D, 1-day dark-imbibed spores; 4D1hB, 1-hr blue light irradiation samples after 4 days of incubation in the dark. The remaining samples are labeled similarly, with R indicating red light.

(B) Relative amounts of expression of five CRY genes and the germination rates in the same samples. Each point represents the average of the amounts of accumulation from two experiments. The symbols indicate the light conditions: circles, dark; squares, red light; triangles, blue light. The amounts of cDNA were determined from densitometric analysis of the gel images by using National Institutes of Health Image software.