The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis.

Adenosine plays an important role in the physiological and pathological conditions of the body by combining different types of adenosine receptors widely distributed in various tissues in the body. In present study, an acute model for paraquat-poisoning in Caenorhabditis elegans was established for quantitative assessment via a time-dose-mortality (TDM) modeling technique with various paraquat doses over 8 hours. Adenosine was first used to precondition at high, medium, and low concentrations and the survival rate of C. elegans was recorded to evaluate adenosine antistress protection against paraquat damage. The results revealed that the TDM model was good for the quantitative assessment of paraquat-poisoning on C. elegans based on the Hosmer-Lemeshow test for homogeneity of modeling ( P = .38). The survival rates of adenosine-preconditioned C. elegans have a dose-dependent association with adenosine concentration. At 3000 μM (high concentration) and 300 μM (medium concentration), adenosine-preconditioned C. elegans still had survival rates of 5.38% ± 1.68% and 5.0% ± 1.19% in the subsequent 8 hours observation period. On the contrary, the survival rates of those receiving 30 μM (low concentration) and the 0 μM (unpreconditioned treatment) were zero. To conclude, adenosine preconditioning had protective effects on C. elegans intoxicated with paraquat by decreasing its mortality rate.

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