Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR

Background: Rust caused by Puccinia psidii Winter has been limiting for the establishment of new Eucalyptus plantations, as well as for resprouting of susceptible genetic materials. Identifying host genes involved in defense responses is important to elucidate resistance mechanisms. Reverse transcription-quantitative PCR is the most common method of mRNA quantitation for gene expression analysis. This method generally employs a reference gene as an internal control to normalize results. A good endogenous control transcript shows minimal variation due to experimental conditions. Findings. We analyzed the expression of 13 genes to identify transcripts with minimal variation in leaves of 60-day-old clonal seedlings of two Eucalyptus clones (rust-resistant and susceptible) subjected to biotic (P. psidii) and abiotic (acibenzolar-S-methyl, ASM) stresses. Conclusions. For tissue samples of clones that did not receive any stimulus, a combination of the eEF2 and EglDH genes was the best control for normalization. When pathogen-inoculated and uninoculated plant samples were compared, eEF2 and UBQ together were more appropriate as normalizers. In ASM-treated and untreated leaves of both clones, transcripts of the CYP and elF4B genes combined were the ones with minimal variation. Finally, when comparing expression in both clones for ASM-treated leaves, P. psidii-inoculated leaves, ASM-treated plus P. psidii-inoculated leaves, and their respective controls, the genes with the most stable expression were EgIDH and UBQ. The chitinase gene, which is highly expressed in studies on plant resistance to phytopathogens, was used to confirm variation in gene expression due to the treatments. © 2010 Laia et al; licensee BioMed Central Ltd.

Does disturbance affect bud bank size and belowground structures diversity in Brazilian subtropical grasslands?

Brazilian Campos grasslands are ecosystems under high frequency of disturbance by grazing and fires. Absence of such disturbances may lead to shrub encroachment and loss of plant diversity. Vegetation regeneration after disturbance in these grasslands occurs mostly by resprouting from belowground structures. We analyzed the importance of bud bank and belowground bud bearing organs in Campos grasslands. We hypothesize that the longer the intervals between disturbances are, the smaller the size of the bud bank is. Additionally, diversity and frequency of belowground organs should also decrease in areas without disturbance for many years. We sampled 20 soil cores from areas under different types of disturbance: grazed, exclusion from disturbance for two, six, 15 and 30 years. Belowground biomass was sorted for different growth forms and types of bud bearing organs. We found a decrease in bud bank size with longer disturbance intervals. Forbs showed the most drastic decrease in bud bank size in the absence of disturbance, which indicates that they are very sensitive to changes in disturbance regimes. Xylopodia (woody gemmiferous belowground organs with hypocotyl-root origin) were typical for areas under influence of recurrent fires. The diversity of belowground bud bearing structures decreased in the absence of disturbance. Longer intervals between disturbance events, resulting in decrease of bud bank size and heterogeneity of belowground organs may lead to the decline and even disappearance of species that relay on resprouting from the bud bank upon disturbance. (C) 2014 Elsevier GmbH. All rights reserved.