Figure 4.

Protonemal Colonies of Cryptochrome Disruptants under Different Light Conditions and Complementation of Cryptochrome Disruptants by Overexpression of PpCRY1b cDNA.

Colonies of wild type, cry1a, cry1b, cry1a cry1b, and PpCRY1b cDNA overexpressors in the cry1a cry1b-1 background were inoculated on agar plates. These plates were incubated under continuous white, blue, or red light for 10 days.

(A), (E), (I), and (M) Wild type (WT).

(B), (F), (J), (N), and (O) cry1a-1.

(C), (G), and (K) cry1b-1.

(D), (H), (L), and (P) cry1a cry1b-1.

(Q) to (T) PpCRY1b cDNA overexpressor in cry1a cry1b-1 (CRY1b+-1/ cry1a cry1b-1).

(U) Different PpCRY1b cDNA overexpressor in cry1a cry1b-1 (CRY1b+-2/cry1a cry1b-1).

The light conditions are as given: white light ([A] to [D] and [Q]); blue light ([E] to [H] and [R]); and red light ([I] to [J] and [S]). The blue light–grown protonemata are shown in (M) to (P) and (T) and (U). The protonema cell walls were stained with 0.1% (v/v) Calcofluor white to help distinguish each cell. The appearance of protonemata of cry1b strains was not distinguished from that of cry1a strain protonemata. Bars in (L) and (S) = 2 mm for (A) to (L) and (Q) to (S); bars in (M) to (P) and (T) and (U) = 200 μm.

(V) Overexpression of PpCRY1b transcripts (arrowhead) in CRY1b+-1/ cry1a cry1b-1 strains was confirmed by RNA gel blot analysis. The asterisk shows the putative transcript derived from the disrupted PpCRY1b locus.