Analysis of mRNA RGFP966 purchase levels after co-cultivation of Trichoderma with Rhizoctonia solani revealed a significantly enhanced expression of Trive160502 (p = 0.000) and Trive180426 (p = 0.031) in T. virens, Triat152366 (p = 0.027) and Triat210209 (p = 0.000) in T. atroviride, and Trire56426 (p = 0.000) in T. reesei upon contact with the host fungus (Figure 3). On the other hand, expression of Triat142946 (p = 0.000), Triat136196 (p = 0.000) in T. atroviride,
Trive92622 (p = 0.000), Trive47976 (p = 0.000), Trive30459 (p = 0.034) in T. virens, and Trire70139 (p = 0.032), Trire119819 (p = 0.000) in T. reesei Vactosertib order was significantly PLX 4720 decreased in the presence of R. solani compared to the corresponding controls. Transcript levels of Triat290043 (p = 0.971), Triat142943 (p = 0.093), and Trire82246 (p = 0.102) were unaffected by the presence of R. solani. Again no transcript could be detected for Triat46847. Expression of Triat46847 was further assessed on both plates and in liquid minimal and full media and under different developmental stages (vegetative growth, conidiation) of the fungus. No transcript could
be detected under all the conditions tested (data not shown). Figure 3 Relative transcription ratios Liothyronine Sodium of PAQR family (class VIII) members. mRNA levels of the respective genes of T. atroviride (A), T. virens (B) and T. reesei (C) upon direct contact with the host fungus R. solani (black bars) were assessed by RT-qPCR and compared to a control where the respective Trichoderma species was grown alone (white bars). Samples of the gene
with highest expression in the control condition were arbitrarily assigned the factor 1. sar1 was used as reference gene. Analysis of the location of the seven PAQR-encoding genes in the genome of T. atroviride revealed that three of them (Triat142946, Triat142943, Triat46847) are in close vicinity on scaffold 19 (Figure 4). This is similar in T. virens and T. reesei for the orthologues of Triat142946 and Triat142943 suggesting the possibility that the third T. atroviride gene (Triat46847), which was found not to be expressed under any of the conditions tested, may have resulted from gene duplication with subsequent inactivation. Figure 4 Schematic drawing of the T. atroviride genomic locus with the PAQR (class VIII)-encoding genes Triat142946, Triat142943, and Triat46847 and the loci with their orthologues in T. virens and T. reesei . Scaffolds and position numbers are given as specified in the respective genome databases [57–59].