Detection of differentially methylated regions of irradiated fig tree selections

Fig tree (Ficus carica L.) breeding programs using conventional methods, such as directed crosses, to obtain new cultivars, are unworkable in many countries, including Brazil. Consequently, genetic breeding through mutagenesis has emerged as an important line of research that can improve this crop, and be a significant source of information about this species and assist in the implementation of propagation projects and appropriate management. The aim of this study was to verify the existence of epigenetic variability attributable to DNA methylation in irradiated fig selections when compared both to each other and to the main commercial cultivar, “Roxo-de-Valinhos”, which had previously used methylation-sensitive amplified polymorphism (MSAP) and DNA sequencing to detect the position of polymorphic regions, analyzable by bioinformatic tools. The sequencing of DNA, isolated from the differentially methylated sites, makes it possible to observe different patterns of methylation by sequencing the treated DNA with sodium bisulfite in the coding regions of regulatory genes active in the development, and fruit ripening stages. Furthermore, they have been found in the mitochondrial DNA of treatments which regulate the supply of energy in Adenosine triphosphate (ATP) form in plants. Closely related to their development, they justify the different phenotypes found in both fruit and plant growth that have suffered stress due to exposure to gamma radiation. Thus, future studies on gene expression in treatments have emerged as an extremely important strategy for understanding these complex regulatory systems, which may lead to the identification of genes of agricultural interest for the fig tree crop, and allow for manipulation and subsequent propagation of improved crops for commercial purposes

Parâmetros biométricos, acúmulo de prolina e identificação de respostas moleculares, por cDNA-AFLP, ao estresse por déficit hídrico em cana-de-açúcar

A cana-de-açúcar (Saccharum spp.) é uma das mais importantes culturas e o Brasil é considerado o maior produtor mundial de etanol. Entretanto, sua produção é diretamente influenciada pelos estresses ambientais, entre eles, o déficit hídrico. Sendo assim, o objetivo do trabalho foi avaliar o comportamento, por meio de análises biométricas e bioquímica de três cultivares de cana-de-açúcar tolerantes (SP83-5073 e RB86-7515) e sensível (SP86-155) ao estresse por déficit hídrico, e verificar a expressão gênica diferencial dependente do genótipo, utilizando a técnica de cDNA-AFLP. Aos 186 dias após o plantio, em casa de vegetação, as cultivares foram submetidas a 1, 3, 5 e 10 dias de supressão da rega. Foi verificado na cultivar RB86-7515 o maior diâmetro do colmo, matéria fresca e seca de folhas e bainha e na cultivar SP86-155, maior altura inicial, número de folhas e matéria fresca de colmo. O teor de prolina livre nos palmitos foi estatisticamente significativo a partir do terceiro dia de supressão da rega, onde foi verificada a maior média na cultivar RB86-7515, com 10 dias. Pela técnica de cDNA-AFLP, foram detectados fragmentos expressos (FEs), identificados pela ausência do fragmento na cultivar sensível e fragmentos exclusivos expressos nas cultivares tolerantes (FEs-T) mas desde que ausentes em suas planta controle. Com 10 combinações de primers seletivos EcoRI/MseI, em gel de poliacrilamida, foram observados 1.077 fragmentos expressos nas cultivares tolerantes controle e sob estresse hídrico. Destes, 463 fragmentos na SP83-5073, sendo 170 fragmentos nas plantas sob estresse hídrico e 18 fragmentos exclusivos. Na RB86-7515, observou-se 614 fragmentos, onde 283 fragmentos nas plantas sob supressão da rega e 30 fragmentos exclusivos. A análise comparativa do perfil de expressão revelou fragmentos de transcritos...Sugarcane (Saccharum spp.) is one of the most important crops and Brazil is the largest world producer of ethanol. However, their production is directly influenced to environmental stresses, as a water stress. So, the aims of this research were to evaluate the behavior of three sugarcane tolerant cultivars (SP83-5073 e RB86-7515) and sensitive (SP86-155) to water stress by biometrical and biochemical analysis, and verified the differential gene expression genotype dependent, using the cDNA-AFLP technique. At 186 days after planting, in greenhouse, cultivars were submitted to 1, 3, 5 and 10 days of water suppression. In cultivar RB86-7515, were observed the largest stalk diameter, leaves and sheath fresh and dried weigh and in cultivar SP86-155, largest initial height, leaves number and stalk fresh mass. Free proline content in sugarcane heart was statistically significative after the 3rd day of water suppression, which the large media was observed in cultivar RB86-7515, with 10 days of water suppression. Through cDNA-AFLP technique, expressed fragments were detected, identified by the fragment absence in sensitive cultivar, and exclusive fragments were expressed in tolerant cultivars, but absent in control plants. Using 10 combinations of selective primers EcoRI/MseI, in poliacrilamide gel, 1,077 expressed fragments in control tolerant cultivars and under water stress were observed. From this total, 463 fragments in SP83-5073, 170 fragments in water-stressed plants and 18 exclusive fragments. In RB86-7515, 614 fragments were observed, 283 fragments in plants under water suppression and 30 exclusive fragments. Profile expression comparative analysis showed fragments of similar transcriptions to sorghum P450 Citocrome, beta-glucosidase, glyceraldehyde-3-phosphate dehydrogenase and DNA Helicase. These... (Complete abstract click electronic access below)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Differences in global gene expression in muscle tissue of Nellore cattle with divergent meat tenderness

Background: Meat tenderness is the consumer's most preferred sensory attribute. This trait is affected by a number of factors, including genotype, age, animal sex, and pre-and post-slaughter management. In view of the high percentage of Zebu genes in the Brazilian cattle population, mainly Nellore cattle, the improvement of meat tenderness is important since the increasing proportion of Zebu genes in the population reduces meat tenderness. However, the measurement of this trait is difficult once it can only be made after animal slaughtering. New technologies such as RNA-Seq have been used to increase our understanding of the genetic processes regulating quantitative traits phenotypes. The objective of this study was to identify differentially expressed genes related to meat tenderness, in Nellore cattle in order to elucidate the genetic factors associated with meat quality. Samples were collected 24 h postmortem and the meat was not aged. Results: We found 40 differentially expressed genes related to meat tenderness, 17 with known functions. Fourteen genes were up-regulated and 3 were down-regulated in the tender meat group. Genes related to ubiquitin metabolism, transport of molecules such as calcium and oxygen, acid-base balance, collagen production, actin, myosin, and fat were identified. The PCP4L1 (Purkinje cell protein 4 like 1) and BoLA-DQB (major histocompatibility complex, class II, DQ beta) genes were validated by qRT-PCR. The results showed relative expression values similar to those obtained by RNA-Seq, with the same direction of expression (i.e., the two techniques revealed higher expression of PCP4L1 in tender meat samples and of BoLA-DQB in tough meat samples). Conclusions: This study revealed the differential expression of genes and functions in Nellore cattle muscle tissue, which may contain potential biomarkers involved in meat tenderness.project "Genomic tools for the genetic improvement of traits of direct economic importance in Nelore cattle"; Sao Paulo Research Foundation - FAPESP [2009/16118-5, 2013/09190-7]Open access journal.This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Expression of genes related to mitochondrial function in Nellore cattle divergently ranked on residual feed intake

Several measures have been proposed to investigate and improve feed efficiency in cattle. One of the most commonly used measure of feed efficiency is residual feed intake (RFI), which is estimated as the difference between actual feed intake and expected feed intake based on the animal's average live weight. This measure permits to identify and select the most efficient animals without selecting for higher mature weight. Mitochondrial function has been indicated as a major factor that influences RFI. The analysis of genes involved in mitochondrial function is therefore an alternative to identify molecular markers associated with higher feed efficiency. This study analyzed the expression of PGC1 alpha, TFAM, UCP2 and UCP3 genes by quantitative real-time PCR in liver and muscle tissues of two groups of Nellore cattle divergently ranked on RFI values in order to evaluate the relationship of these genes with RFI. In liver tissue, higher expression of TFAM and UCP2 genes was observed in the negative RFI group. Expression of PGC1 alpha gene did not differ significantly between the two groups, whereas UCP3 gene was not expressed in liver tissue. In muscle tissue, higher expression of TFAM gene was observed in the positive RFI group. Expression of PGC1 alpha, UCP2 and UCP3 genes did not differ significantly between the two groups. These results suggest the use of TFAM and UCP2 as possible candidate gene markers in breeding programs designed to increase the feed efficiency of Nellore cattle.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP