Validation of reference genes for RT-qPCR normalization in common bean during biotic and abiotic stresses

Selection of reference genes is an essential consideration to increase the precision and quality of relative expression analysis by the quantitative RT-PCR method. The stability of eight expressed sequence tags was evaluated to define potential reference genes to study the differential expression of common bean target genes under biotic (incompatible interaction between common bean and fungus Colletotrichum lindemuthianum) and abiotic (drought; salinity; cold temperature) stresses. The efficiency of amplification curves and quantification cycle (C (q)) were determined using LinRegPCR software. The stability of the candidate reference genes was obtained using geNorm and NormFinder software, whereas the normalization of differential expression of target genes [beta-1,3-glucanase 1 (BG1) gene for biotic stress and dehydration responsive element binding (DREB) gene for abiotic stress] was defined by REST software. High stability was obtained for insulin degrading enzyme (IDE), actin-11 (Act11), unknown 1 (Ukn1) and unknown 2 (Ukn2) genes during biotic stress, and for SKP1/ASK-interacting protein 16 (Skip16), Act11, Tubulin beta-8 (beta-Tub8) and Unk1 genes under abiotic stresses. However, IDE and Act11 were indicated as the best combination of reference genes for biotic stress analysis, whereas the Skip16 and Act11 genes were the best combination to study abiotic stress. These genes should be useful in the normalization of gene expression by RT-PCR analysis in common bean, the most important edible legume.

Tidal perturbations of the rotation of the moon: the creep tide approach

In this communication we report results from the application to the study of the rotation of the Moon of the creeping tide theory just proposed (Ferraz-Mello, Cel. Mech. Dyn. Astron., submitted. ArXiv astro-ph 1204.3957). The choice of the Moon for the first application of this new theory is motivated by the fact that the Moon is one of the best observed celestial bodies and the comparison of the theoretical predictions of the theory with observations i may validate the theory or point out the need of further improvements. Particularly, the tidal perturbations of the rotation of the Moon - the physical libration of the Moon - have been detected in the Lunar Laser Ranging measurements (Williams et al. JGR 106, 27933, 2001). The major difficulty in this application comes from the fact that tidal torques in a planet-satellite system are very sensitive to the distance between the two-bodies, which is strongly affected by Solar perturbations. In the case of the Moon, the main solar perturbations - the Evection and the Variation - are more important than most of the Keplerian oscillations, being smaller only than the first Keplerian harmonic (equation of the centre). Besides, two of the three components of the Moon's libration in longitude whose tidal contributions were determined by LLR are related to these perturbations. The results may allow us to determine the main parameter of a possible Moon's creeping tide. The preliminary results point to a relaxation factor (gamma) 2 to 4 times smaller than the one predicted from the often cited values of thr Moon's quality factor Q (between 30 and 40), and points to larger Q values.