RESUMO
In the original publication the Figs. 3 and 4 are published inappropriately. The correct version of Figs. 3 and 4 is provided in this correction.
RESUMO
An obligate halophile fungal was isolated from 275 m deep marine sediments and is characterized here for the first time. Its optimal growth was at 15% NaCl even though it was able to grow at 25% and is incapable of growth with no NaCl. Based on its morphological characteristics as conidia chain production in a single phialide, the fungal is related to the genus Aspergillus, subgenus Polypaecilum. Phylogenetic molecular analysis using several markers (ITS1-2; RPB1; RPB2; Cct8; TSR1; CaM; BenA) places the fungal isolate closer to Aspergillus salinarus and A. baarnensis. However, its morphological and molecular differences establish it as a new species, Aspergillus loretoensis sp. nov.
Assuntos
Aspergillus/genética , Tolerância ao Sal , Aspergillus/classificação , Aspergillus/isolamento & purificação , Aspergillus/metabolismo , Baías/microbiologia , Genes Fúngicos , Sedimentos Geológicos/microbiologia , FilogeniaRESUMO
The red abalone Haliotis rufescens is one of the most important species for aquaculture in Baja California, México, and despite this, few gene expression studies have been done in tissues such as gill, head and gonad. For this purpose, reverse transcription and quantitative real time PCR (RT-qPCR) is a powerful tool for gene expression evaluation. For a reliable analysis, however, it is necessary to select and validate housekeeping genes that allow proper transcription quantification. Stability of nine housekeeping genes (ACTB, BGLU, TUBB, CY, GAPDH, HPRTI, RPL5, SDHA and UBC) was evaluated in different tissues of red abalone (gill, head and gonad/digestive gland). Four-fold serial dilutions of cDNA (from 25 ngµL(-1) to 0.39 ngµL(-1)) were used to prepare the standard curve, and it showed gene efficiencies between 0.95 and 0.99, with R(2)=0.99. geNorm and NormFinder analysis showed that RPL5 and CY were the most stable genes considering all tissues, whereas in gill HPRTI and BGLU were most stable. In gonad/digestive gland, RPL5 and TUBB were the most stable genes with geNorm, while SDHA and HPRTI were the best using NormFinder. Similarly, in head the best genes were RPL5 and UBC with geNorm, and GAPDH and CY with NormFinder. The technical variability analysis with RPL5 and abalone gonad/digestive gland tissue indicated a high repeatability with a variation coefficient within groups ≤ 0.56% and between groups ≤ 1.89%. These results will help us for further research in reproduction, thermoregulation and endocrinology in red abalone.
Assuntos
Perfilação da Expressão Gênica/normas , Genes Essenciais , Moluscos/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Animais , Feminino , Perfilação da Expressão Gênica/métodos , Gônadas/metabolismo , Masculino , Moluscos/metabolismo , Controle de Qualidade , Reação em Cadeia da Polimerase em Tempo Real/métodos , Padrões de Referência , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas , Estudos de Validação como AssuntoRESUMO
Oocytes of Crassostrea gigas and Mytilus galloprovincialis are arrested in metaphase I when they are spawned and ready to be fertilized. To investigate the role of MAP kinase in maintaining metaphase I arrest, oocytes were exposed to the MEK inhibitor U0126, and the effects on chromosome behavior and MAPK activity were examined by bisbenzimide staining and in immunoblots with anti-phospho MAPK antibodies. Following treatment with 50 microM U0126, active MAPK was undetectable and oocytes resumed meiosis, forming enlarged polar bodies and undergoing chromosome decondensation. Prophase stage oyster oocytes maturing spontaneously in seawater completed germinal vesicle breakdown in the presence of U0126, but failed to arrest in metaphase I, and also formed polar bodies and underwent chromosome decondensation. Treatment of oyster oocytes with the protein synthesis inhibitor, emetine (500 microM), also caused them to resume meiosis, although substantial MAPK activity remained. Levels of phospho-MEK also decreased during emetine treatment. 35 S-methionine incorporation in emetine treated oocytes was reduced to only 5% of control values. These data show that, while active MAPK is necessary to maintain metaphase I arrest, other proteins are also required.