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Reverse transcription-quantitative PCR (RT-qPCR) is an analytical tool for gene expression quantification. Reference genes are not yet available for gene expression analysis during interactions of Ralstonia solanacearum with ‘Hawaii 7996’ (the most stable source of resistance in tomato). Here, we carried out a multi-algorithm stability analysis of eight candidate reference genes during interactions of ‘Hawaii 7996’ with one incompatible/avirulent and two compatible/virulent (= resistance-breaking) bacterial isolates. Samples were taken at 24- and 96-h post-inoculation (HPI). Analyses were performed using the ∆∆Ct method and expression stability was estimated using BestKeeper, NormFinder, and geNorm algorithms. TIP41 and EF1α (with geNorm), TIP41 and ACT (with NormFinder), and UBI3 and TIP41 (with BestKeeper), were the best combinations for mRNA normalization in incompatible interactions at 24 HPI and 96 HPI. The most stable genes in global compatible and incompatible interactions at 24 HPI and 96 HPI were PDS and TIP41 (with geNorm), TIP41 and ACT (with NormFinder), and UBI3 and PDS/EXP (with BestKeeper). Global analyses on the basis of the three algorithms across 20 R. solanacearum-tomato experimental conditions identified UBI3, TIP41 and ACT as the best choices as reference tomato genes in this important pathosystem.

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Teak is a forest species that has assumed great importance in Brazil, where it has found excellent conditions for development since its introduction into the country in the 1960s. However, phytosanitary problems are beginning to threaten the production of this timber species. An example is teak canker, caused by the fungus Lasiodiplodia theobromae (Lt), which has only recently been reported in Brazil, and for which, therefore, there are no recommended control methods. Thus, this study evaluated the control of this pathogen, investigating the potential of the biocontrol agents (BCAs) Trichoderma spp., Bacillus sp. and Enterobacter sp., initially through in vitro assays and, subsequently, with in vivo tests. According to the in vitro assay results, the Trichoderma isolates CEN162 and CEN1153 and the strain of Bacillus sp. (UnB1366) were the treatments that stood out, as they were able to completely inhibit mycelial growth of some isolates of Lt. When these isolates were tested in a preventive way, the control levels varied depending on the Lt isolate and the antagonist-clone interaction, where CEN162 (T. asperellum) and UnB166 (Bacillus sp.) showed 100% control. Thus, there is a positive correlation between the in vitro and in vivo tests, since the same BCAs stood out. Although good levels of control have been obtained with the BCAs used, it can be concluded that there is a variation in the antagonism to different Lt isolates or even in the antagonist-clone interaction, corroborating the information available in the scientific literature on this plant-pathogenic fungus.