SpringerImages - Expression of brlA(p/l)::lacZ, abaA(p/l)::lacZ, and medA(p/l)::lacZ in the wild-type strain and ΔchsA ΔchsC double mutant. a Messenger RNA levels of brlA,abaA,medA, and lacZ. Total RNA was extracted from strains cultured for 0 or 24 h on solid MMG after being transferred from liquid MMG. Strains used are ///pJA22-1 (wild type containing brlA(p/l)::lacZ), ΔAC/pJA22-2 (ΔchsA ΔchsC mutant containing brlA(p/l)::lacZ), ///pABAL-8 (wild type containing abaA (p/l)::lacZ), ΔAC/pABAL-6 (ΔchsA ΔchsC mutant containing abaA (p/l)::lacZ), ///pML-2 (wild type containing medA(p/l)::lacZ), and ΔAC/pML-3 (ΔchsA ΔchsC mutant containing medA(p/l)::lacZ). Ribosomal RNA (rRNA) stained with ethidium bromide is shown as a loading control. b–e In-situ staining. ///pJA22-1 (bupper panel), ΔAC/pJA22-2 (blower panel), ///pABAL-8 (cupper panel), ΔAC/pABAL-6 (clower panels), ///pML-2 (dupper panel), ΔAC/pML-3 (dlower panel), ///-10 (wild type, eupper panel), and ΔAC/A-4 (ΔchsA ΔchsC mutant, elower panel) were grown on solid MMG for 24 h and subjected to in-situ staining as described in the text

Caption

Fig 3

Expression of brlA(p/l)::lacZ, abaA(p/l)::lacZ, and medA(p/l)::lacZ in the wild-type strain and ΔchsA ΔchsC double mutant. a Messenger RNA levels of brlA,abaA,medA, and lacZ. Total RNA was extracted from strains cultured for 0 or 24 h on solid MMG after being transferred from liquid MMG. Strains used are ///pJA22-1 (wild type containing brlA(p/l)::lacZ), ΔAC/pJA22-2 (ΔchsA ΔchsC mutant containing brlA(p/l)::lacZ), ///pABAL-8 (wild type containing abaA (p/l)::lacZ), ΔAC/pABAL-6 (ΔchsA ΔchsC mutant containing abaA (p/l)::lacZ), ///pML-2 (wild type containing medA(p/l)::lacZ), and ΔAC/pML-3 (ΔchsA ΔchsC mutant containing medA(p/l)::lacZ). Ribosomal RNA (rRNA) stained with ethidium bromide is shown as a loading control. b–e In-situ staining. ///pJA22-1 (bupper panel), ΔAC/pJA22-2 (blower panel), ///pABAL-8 (cupper panel), ΔAC/pABAL-6 (clower panels), ///pML-2 (dupper panel), ΔAC/pML-3 (dlower panel), ///-10 (wild type, eupper panel), and ΔAC/A-4 (ΔchsA ΔchsC mutant, elower panel) were grown on solid MMG for 24 h and subjected to in-situ staining as described in the text

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Northern analysis confirmed the similarity of expressions of these fusion genes to those of brlA , abaA , and medA (Fig. 3 a).

In in-situ staining, expressions of brlA (p/l): lacZ and abaA (p/l):: lacZ were detected in morphologically aberrant sterigmata and conidiophore vesicles of the ΔAC mutants (Fig. 3 b, c).

Regarding the ΔAC mutants with abaA (p/l):: lacZ , we readily found poorly stained conidiophores that had developed long chains of aberrant sterigmata in the ΔAC mutant (Fig. 3 c, left).

Expression of medA (p/l):: lacZ was observed in both hyphae and conidiophores in either the wild-type or ΔAC mutant strains (Fig. 3 d).

The results of in-situ staining (Fig. 3 ) suggest that induction of abaA expression does not occur during reiteration of abnormal sterigmata, while brlA and medA are expressed.

Northern analysis (Fig. 2 ) and in-situ staining (Fig. 3 ) revealed that medA expression in the ΔAC mutants was not disturbed.

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