Figure 1.
Blue-light perception and signaling regulate H. jecorina sexual development.
(A) Schematic representation of the mating assay. Two single-ascospore cultures (#1 & #2) were inoculated on a 10-cm MEA plate as indicated. If sexual development occurs, stromata will be found at the interaction zone. (B–D) Sexual development of wild-type CBS999.97(1–1)×CBS999.97(1–2) strain under a 24L, 12L12D, or 24D photoperiod. (E–G) Sexual development of Δblr1, Δblr2, Δblr1 Δblr2 (Δblr1,2) and Δenv1 diploid mutants under a 24-h, 12-h, or 0-h photoperiod. (H) Sexual development of crossing Δblr1 Δenv1 (1–1x1–2) under constant illumination. (I) Sexual development of crossing Δenv1(1–1) x Δblr1 Δenv1(1–2) under constant illumination.
Figure 2.
Light affects perithecia development.
CBS999.97(1–1)×CBS999.97(1–2) crossing were carried out under a 12L12D photoperiod for 12 days (A) or in constant darkness for 7 days (B, C) or 14 days (D), respectively. Frozen sections of stromata were visualized by hematoxylin and eosin stain. (White scale bars: 50 mm).
Figure 3.
Stroma induction by conidia in a 12L12D photoperiod.
Female recipient strains grew on a 10-cm MEA plate under a 12L12D photoperiod for 7 days, including CBS999.97(1–1), CBS999.97(1–2) (A), QM6a (B), and CBS999.97(1–1) Δenv1, and CBS999.97(1–2) Δenv1 mutants (C, D). The conidia from male strains were spotted onto an indicated white oval region of the female recipient MEA plate. The MEA plate was incubated under a 12L12D photoperiod for 10–14 days. Caffeine (2 mM) was added to the MEA plate in (D). The stromata were marked as indicated.
Figure 4.
Stroma induction by conidia in constant darkness.
Female recipient strains grew on a 10-cm MEA plate under a 12L12D photoperiod for 7 days, including CBS999.97(1–1), CBS999.97(1–2) (A), QM6a (B), and CBS999.97(1–1) Δenv1, and CBS999.97(1–2) Δenv1 mutants (C). The conidia from a male strain were spotted onto an indicated white oval region of a female recipient MEA plate. The MEA plate was incubated under a 24D photoperiod (constant darkness) for 10–14 days. The stromata were marked as indicated.
Figure 5.
Transcription levels of putative genes involved in conidiation and sexual potency.
Total RNAs were extracted from 8 different experimental conditions as indicated. The quality of extracted RNA samples was further analyzed with the RNA 6000 Nano kit by Agilent 2100 Bioanalyzer (see Materials and Methods). (A) Northern blots analysis of act1 (actin) and env1 transcription. The denaturing RNA agarose gel was stained with ethidium bromide, the 18S rRNA and 28S rRNA bands were clearly visible in the intact RNA samples. (B–H) qRT-PCR. Relative transcript abundance of representative genes in sexually potent and impotent conditions. Data were given as relative quantitative (RQ) values to one of the eight conditions as indicated. The transcripts of the ribosome protein gene rpl6e were used for normalization of the qRT-PCR data [43].
Figure 6.
Regulatory targets of sexual potency in the CBS999.97(1–2) wild-type strain (W) and the CBS999.97(1–2) Δenv1 mutant (E).
VENN diagram of genes 2-fold downregulated genes (A) and 2-fold up-regulated genes (B) in the four sexually potent conditions (W-24D, W-12L12D, W-12D12L, E24D) in comparison with the four sexually impotent conditions (E24L, E-12L12D, E-12D12L, W-24L). For details on gene regulation see supplementary files (Tables S1 and S2).
Table 1.
Annotation of 15 representative genes that are transcriptionally down-regulated in the four sexually potent conditions.
Table 2.
Relative transcriptional levels of 15 representative down-regulated genes in the 4 sexually potent conditions v.s. the 4 sexually impotent condition.
Table 3.
Annotation of representative genes that are transcriptionally up-regulated in the four sexually potent conditions.
Figure 7.
The effects of Δhpp1 deletion on stroma induction under a 12L12D or 24L photoperiod.
Sexual development of the Δhpp1 mutant was determined as described in Figure 1.