Validación de nuevos genes de referencia para estudios de expresión diferencial de genes involucrados en la síntesis de antocianinas en lechuga (Lactuca sativa L.) y especies silvestres relacionadas

Publishe

Selection of Novel Reference Genes by RNA-Seq and Their Evaluation for Normalising Real-Time qPCR Expression Data of Anthocyanin-Related Genes in Lettuce and Wild Relatives

Lettuce is a popular vegetable source of bioactive compounds, like anthocyanins, powerful antioxidants present in red and semi-red varieties. Selection of reliable reference genes (RGs) for the normalization of real-time quantitative PCR (qPCR) data is crucial to obtain accurate gene expression results. Among the genes with totally unrelated biological functions, six candidate RGs (ADF2, CYB5, iPGAM, SCL13, TRXL3-3, and VHA-H) with low variation in expression according to RNA-seq analyses, were selected for future expression studies of anthocyanin-related genes in three different experiments: leaf colour comparison (green vs. red) in commercial varieties; tissue comparison (leaf vs. stem) in a wild relative; and drought stress experiment in commercial and traditional varieties, and a wild relative. Expression profiles of the candidate RGs were obtained by qPCR and their stability was assessed by four different analytical tools, geNorm, NormFinder, BestKeeper, and Delta Ct method, all integrated in RefFinder. All results considered, we recommend CYB5 to be used as RG for the leaf colour experiment and TRXL3-3 for the tissue and drought stress ones, as they were the most stable genes in each case. RNA-seq is useful to preselect novel RGs although validation by qPCR is still advisable. These results provide helpful information for gene expression studies in Lactuca spp. under the described conditions.Publishe

Determinación de capsicinoides en germoplasma del género Capsicum spp

Publishe

Estudios sobre el carácter picante del pimiento

1 .pdf (1 Pag.) copia de la noticia.Este proyecto es un buen ejemplo de colaboración entre distintos organismos de investigación aragoneses.Peer reviewe

Molecular characterization of spanish onion (Allium cepa L.) landraces and related allium using microsatellite markers

EUCARPIA. European Association for Research on Plant Breedin

Allelic variation in a putative gene related to pungency in pepper (Capsicum spp.)

This study was supported by the Spanish Ministry of Science (project INIA RTA2008-00095-00-00)and by the Aragón Gobernment (Group A16

Análisis del perfil de capsicinoides en germoplasma de Capsicum spp.

2 Pags.- 1 Fig.En este trabajo se ha llevado a cabo la identificación de capsicinoides en un conjunto de genotipos de especies cultivadas y silvestres de pimiento, mediante un método de HPLC-MS/MS y se han analizado los distintos perfiles de capsicinoides en el género Capsicum. Además, se ha realizado una caracterización organoléptica del carácter picante en los genotipos analizados.Estudio financiado por el Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, INIA (RTA2008-00095-00-00) y el Gobierno de Aragón (grupos A16, A03 y E39)Peer reviewe

Inheritance of capsaicin and dihydrocapsaicin, determined by HPLC-ESI/MS, in an intraspecific cross of Capsicum annuum L.

The quantitative inheritance of capsaicin and dihydrocapsaicin contents in fruits has been studied in an intraspecific cross of Capsicum annuum L. across two different environments, namely, fruits developed in spring and summer. A liquid chromatography-electrospray ionization/time-of-flight mass spectrometry [HPLC-ESI/MS(TOF)] method was used to identify and quantify capsaicin and dihydrocapsaicin in extracts of pepper fruits. The analytical method used was able to determine the pungency of genotypes that, using other methods, would have been classified as non-pungent. Capsaicin and dihydrocapsaicin contents varied largely among families, and families did not respond similarly in producing these capsaicinoids when their fruits were grown in spring and summer, with some families showing no increase, whereas in others, the increase was more than 2-fold. Heterosis for the pungency trait, assessed by the capsaicin and dihydrocapsaicin contents in fruits, was found, indicating the existence of epistasis, over-dominance, or dominance complementation. Non-pungent parent alleles contributed to the capsaicin and dihydrocapsaicin contents since transgressive segregation did occur. Furthermore, the type of gene action varied between capsaicin and dihydrocapsaicin, and a seasonal effect during fruit development could affect gene action. © 2007 American Chemical Society.Peer Reviewe

Determination of capsaicin and dihydrocapsaicin in Capsicum fruits by liquid chromatography-electrospray/time-of-flight mass spectrometry

A simple, highly selective, sensitive, and reproducible liquid chromatography-electrospray ionization/ time-of-flight mass spectrometry method has been developed for the direct and simultaneous determination of capsaicin and dihydrocapsaicin in Capsicum fruit extracts. Capsaicin and dihydrocapsaicin are the two major members of the so-called capsaicinoid family, which includes other minor analogues, and usually account for at least 90% of the pungency trait in Capsicum fruits. Chromatographic separation of capsaicin and dihydrocapsaicin was achieved with a reversed-phase chromatography column, using a gradient of methanol and water. Quantification was done using as an internal standard (4,5-dimethoxybenzyl)-4-methyloctamide, a synthetic capsaicin analogue not found in nature. Analytes were base-peak resolved in less than 16 min, and limits of detection were 20 pmol for capsaicin and 4 pmol for dihydrocapsaicin. The intraday repeatability values were lower than 0.5 and 12% for retention time and peak area, respectively, whereas the interday repeatability values were lower than 0.6 and 14% for retention time and peak area, respectively. Analyte recoveries found were 86 and 93% for capsaicin and dihydrocapsaicin, respectively. The method developed has been applied to the identification and quantification of capsaicin and dihydrocapsaicin in fruit extracts from different Capsicum genotypes, and concentrations found ranged from 2 to 6639 mg kg-1. © 2006 American Chemical Society.Peer Reviewe