A functional genomics project has been initiated to approach the molecular characterization of the main biological and agronomical traits of citrus. As a key part of this project, a citrus EST collection has been generated from 25 cDNA libraries covering different tissues, developmental stages and stress conditions. The collection includes a total of 22,635 high-quality ESTs, grouped in 11,836 putative unigenes, which represent at least one third of the estimated number of genes in the citrus genome. Functional annotation of unigenes which have Arabidopsis orthologues (68% of all unigenes) revealed gene representation in every major functional category, suggesting that a genome-wide EST collection was obtained. A Citrus clementina Hort. ex Tan. cv. Clemenules genomic library, that will contribute to further characterization of relevant genes, has also been constructed. To initiate the analysis of citrus transcriptome, we have developed a cDNA microarray containing 12,672 probes corresponding to 6875 putative unigenes of the collection. Technical characterization of the microarray showed high intra- and inter-array reproducibility, as well as a good range of sensitivity. We have also validated gene expression data achieved with this microarray through an independent technique such as RNA gel blot analysis.

No abstract

Switchgrass (Panicum virgatum) has received a lot of attention as a forage and bioenergy crop during the past few years. Gene expression studies are in progress to improve new traits and develop new cultivars. Quantitative real time PCR (qRT-PCR) has emerged as an important technique to study gene expression analysis. For accurate and reliable results, normalization of data with reference genes is essential. In this work, we evaluate the stability of expression of genes to use as reference for qRT-PCR in the grass P. virgatum. Eleven candidate reference genes, including eEF-1α, UBQ6, ACT12, TUB6, eIF-4a, GAPDH, SAMDC, TUA6, CYP5, U2AF, and FTSH4, were validated for qRT-PCR normalization in different plant tissues and under different stress conditions. The expression stability of these genes was verified by the use of two distinct algorithms, geNorm and NormFinder. Differences were observed after comparison of the ranking of the candidate reference genes identified by both programs but eEF-1α, eIF-4a, CYP5 and U2AF are ranked as the most stable genes in the samples sets under study. Both programs discard the use of SAMDC and TUA6 for normalization. Validation of the reference genes proposed by geNorm and NormFinder were performed by normalization of transcript abundance of a group of target genes in different samples. Results show similar expression patterns when the best reference genes selected by both programs were used but differences were detected in the transcript abundance of the target genes. Based on the above research, we recommend the use of different statistical algorithms to identify the best reference genes for expression data normalization. The best genes selected in this study will help to improve the quality of gene expression data in a wide variety of samples in switchgrass.

To identify genes associated with the cytokinin-induced enhanced drought tolerance, we analyzed the transcriptome of wild-type and transgenic tobacco (Nicotiana tabacum 'SR1') plants expressing P(SARK)::IPT (for senescence-associated receptor kinase::isopentenyltransferase) grown under well-watered and prolonged water deficit conditions using the tomato GeneChip. During water deficit, the expression of genes encoding components of the carotenoid pathway leading to ABA biosynthesis was enhanced in the wild-type plants, but repressed in the transgenic plants. On the other hand, transgenic plants displayed higher transcript abundance of genes involved in the brassinosteroid biosynthetic pathways. Several genes coding for proteins associated with Chl synthesis, light reactions, the Calvin-Benson cycle and photorespiration were induced in the transgenic plants. Notably, increased transcript abundance of genes associated with PSII, the cytochrome b(6)/f complex, PSI, NADH oxidoreductase and the ATP complex was found in the P(SARK)::IPT plants. The increased transcript abundance was assessed by quantitative PCR and the increased protein levels were confirmed by Western blots. Our results indicated that while the photosynthetic apparatus in the wild-type plants was degraded, photosynthesis in the transgenic plants was not affected and photosynthetic proteins were not degraded. During water deficit, wild-type plants displayed a significant reduction in electron transfer and photochemical quenching, with a marked increase in non-photochemical quenching, suggesting a decrease in energy transfer to the PSII core complexes and an increase in cyclic electron transfer reactions.

No abstract

The epiphytic lichen Ramalina farinacea is distributed throughout the northern hemisphere in which the same two algal Trebouxia species (provisionally named TR1 and TR9) coexist in every thallus. Ramalina farinacea symbionts were characterized based on the two fungal nuclear loci (nrITS and rpb2) along with the primary and secondary structures of nrITS from each Trebouxia species in the Iberian Peninsula and Canary Islands. The results indicated a noticeable genetic differentiation between mycobionts from these two geographic areas and also suggested concerted changes in the three partners of a lichen symbiosis toward two clearly distinguishable 'holobiont' lineages. Modeling of ITS2 RNA secondary structures suggested their temperature sensitivity in TR1 but not in TR9, which was consistent with the observed superior physiological performance of TR9 phycobionts under relatively high temperatures. Both TR1 and TR9 phycobionts have been also found in a variety of taxonomically distinct lichens with a preferably Mediterranean distribution, being TR1 much more widespread than TR9. Our observations support a model in which ecological diversification and speciation of lichen symbionts in different habitats could include a transient phase consisting of associations with more than one photobiont in individual thalli. Such diversification is likely to be promoted by different physiological backgrounds.

No abstract

Elsevier Payri, R.; Salvador Rubio, FJ.; Gimeno, J.; Bracho Leon, G. (2011) es (R. Payri). AbstractThe current work focuses on the study of the influence of pressure and temperature on three important thermodynamic properties: speed of sound, compressibility and density of common diesel fuels. The sample fuels studied were conventional diesel (Repsol Elite), Rape Methyl Ester (typical bio-diesel used in Spain) and a fuel used especially for winter season (Arctic). , FUEL 2011, Vol 90, pp. 1172-1180, doi 10.1016/j.fuel.2010.11.015 measurements at these extreme conditions. A detailed description of the method is presented; also, the obtained results are listed, with an uncertainty of ~0.3%. Density measurements have been performed over a broad range of temperatures (298< T/K < 343), using two commercial devices, obtaining an overall uncertainty of ~0.6%.