RESUMO
As a proven tool, DNA barcoding can identify species rapidly and unambiguously. In this study, we used mtDNA cyt b, COI, and 16s rRNA sequences of six species of Pseudohynobius, Protohynobius puxiongensis, Liua shihi, Ranodon sibiricus, and Pachyhynobius shangchengensis, to reconstruct the phylogenetic relationships using Bayesian inference and maximum likelihood methods. Approximate lineage divergence times were also estimated, the divergence between them was calculated to have taken place mainly in Miocene. Our results showed that: 1) Ps. guizhouensis is an independent and valid species that is a sister species to Ps. kuankuoshuiensis; 2) five Pseudohynobius species formed a monophyletic group; 3) Ps. tsinpaensis is different from L. shihi, and should be classified as belonging to the Liua genus; and 4) Pr. puxiongensis is the sister lineage to all Pseudohynobius species, and should therefore be named Pseudohynobius puxiongensis.
Assuntos
Urodelos/genética , Animais , Teorema de Bayes , Citocromos b/genética , Código de Barras de DNA Taxonômico/métodos , DNA Mitocondrial/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
The evolutionary significant units (ESUs) of the salamander Pachyhynobius shangchengensis (Hynobiidae) in the Dabieshan mountains, southeastern China, were identified based on mitochondrial DNA data. We used methods for detecting cryptic species, such as the minimum spanning tree, the automatic barcode gap discovery, and the generalized mixed Yule-coalescent model; geographical partitioning was also used to identify the ESUs. A total of four ESUs were identified.
Assuntos
DNA Mitocondrial , Evolução Molecular , Urodelos/genética , Animais , Evolução Biológica , China , Genes Mitocondriais , Haplótipos , Filogenia , Filogeografia , Urodelos/classificaçãoRESUMO
The effects of exogenous 5-aminolevulinic acid (ALA) on photosynthesis, plant growth, and the expression of two aquaporin genes in tomato seedlings under control and salinity conditions were investigated. Exogenous ALA application significantly improved net photosynthetic rate (Pn), total chlorophyll content, and plant biomass accumulation of tomato seedlings under salinity stress. As revealed by real-time PCR analyses, after treatment with ALA alone, expression of both LePIP1 and LePIP2 in the two tomato cultivars was up-regulated at 2 h and subsequently decreased to normal levels. Under salinity stress, transcript levels of LePIP1 in both leaves and roots of salt-sensitive cultivars (cv. Zhongza No.9) increased significantly and were considerably higher than in cultivars exposed to ALA alone. In contrast, the expression levels of LePIP1 and LePIP2 in cvs. Jinpeng No.1 cultivars were slightly lower under salinity stress than under ALA treatment. In addition, transcript levels of both LePIP1 and LePIP2 in the roots of Jinpeng No. 1 cultivars were considerably lower than those in the roots of Zhongza No. 9 cultivars under salinity stress, regardless of ALA supplementation, implying that Jinpeng No. 1 cultivars had a better capacity to maintain membrane intrinsic protein stability. Further, ALA application distinctly counteracted the up- or down-regulation of LePIP1 and LePIP2 in both cultivars under salinity stress, in accordance with the improvements instomatal conductance, transpiration rate, and Pn of tomato leaves. The results presented here indicate that ALA controls aquaporin expression, thus, presumably ALA regulates water homeostasis and enhances salt tolerance of tomato seedlings.
Assuntos
Fotossíntese/efeitos dos fármacos , Tolerância ao Sal/efeitos dos fármacos , Plântula/genética , Solanum lycopersicum/crescimento & desenvolvimento , Ácido Aminolevulínico/farmacologia , Clorofila/metabolismo , Solanum lycopersicum/efeitos dos fármacos , Fotossíntese/genética , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Salinidade , Plântula/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacosRESUMO
The objectives of this study were to evaluate protein thermal stability, water-protein interaction, microstructure, and protein conformation between PSE-like and normal chicken breast meat batters. Sixty pale, soft, and exudative (PSE)-like (L*>53, pH24 h<5.7) and 60 normal (46Assuntos
Carne/análise
, Músculos Peitorais/fisiologia
, Conformação Proteica
, Animais
, Galinhas/fisiologia
, Eletroforese em Gel de Poliacrilamida
, Manipulação de Alimentos
, Temperatura Alta
, Desnaturação Proteica
RESUMO
Bone morphogenetic proteins (BMPs) are the key factors in maintaining cell growth and differentiation in ovaries. BMPs initiate signaling by assembling BMP receptors and activating Smads, which in turn alter the expression of target genes. However, little is known about the effect of the deletion of the Bone morphogenetic protein receptor type IB (BMPRIB) on porcine granulosa cell (GCs). The objective of this study was to determine the effects of BMPRIB gene silencing, by small interfering RNA (siRNA), on the apoptosis and steroidogenesis of porcine GCs, and the expression of cell cycle-related and apoptosis-related genes. Results indicate that the BMPRIB siRNA caused specific inhibition of BMPRIB mRNA expression after transfection. Knockdown of the BMPRIB gene significantly inhibited porcine GCs proliferation and estradiol production, while inducing apoptosis of porcine GCs. Additionally, the declined expression of the BMPRIB gene changed the expressions of CylinD2, Cdk2, Bcl-2, and Cyp19a1. These findings provide an important role of BMPRIB in the regulation of apoptosis and steroidogenesis of porcine GCs.
Assuntos
Apoptose/genética , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/genética , Inativação Gênica , Células da Granulosa/metabolismo , RNA Interferente Pequeno/metabolismo , Esteroides/biossíntese , Animais , Tamanho Corporal , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Proliferação de Células , Feminino , Células da Granulosa/citologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Proteína Smad4/genética , Proteína Smad4/metabolismo , Sus scrofa , TransfecçãoRESUMO
In this study, we isolated microsatellite DNA from the Huoyan goose genome with magnetic beads. As a result, 150 positive clones were identified, and 148 microsatellites were found. Among the 148 microsatellites, 69.6% were perfect, 17.6% were imperfect, and the rest were compound type (12.8%). Twenty microsatellite primers were used to screen 90 individuals from 3 Huoyan goose populations. Eight loci were polymorphic with a low number of alleles (2 to 4). The observed and expected heterozygosities ranged from 0.3556 to 1 and from 0.2923 to 0.6868, respectively. All the 8 polymorphic loci were in Hardy-Weinberg equilibrium. These molecular markers will be useful for future studies on population genetic structure and conservation genetics in Huoyan geese.
Assuntos
Gansos/genética , Repetições de Microssatélites , Alelos , Animais , Loci Gênicos , Variação Genética , Genética Populacional , Genótipo , Dados de Sequência MolecularRESUMO
Aquaporins play a direct role in plant water relation under salt stress, but the effects of 5-aminolevulinic acid (ALA) on aquaporin gene expression in salt-treated plants remain unknown. This study investigated the potential effects of exogenous ALA (50 mg/dm3) on aquaporin expression levels under salt stress (75 mM NaCl) in the salt-sensitive (Jinchun No.4) and the relatively salt-tolerant cucumber (Jinyou No.1) seedlings. The expressions of cucumber PIP aquaporin gene (CsPIP1:1) and cucumber NIP aquaporin gene (CsNIP) were analyzed in 20-day-old seedling leaves at 2, 4, 8, 16, and 24 h after ALA treatment. After treatment with saline alone and ALA alone, CsPIP1:1 and CsNIP gene expression levels in the 2 cucumber cultivars increased to maximum at 2 h. The aquaporin gene expression in salt-treated cucumber seedling leaves was considerably higher than that in leaves subjected to exogenous ALA. Further, the aquaporin expression levels in Jinchun No.4 were higher than those in Jinyou No.1, reaching 5.20- and 2-fold induction levels, respectively. After treatment with both ALA and NaCl, the CsNIP gene expression was downregulated in both the cucumber cultivars, while that of CsPIP1:1 decreased at 2 h and then increased to 3.8-fold in Jinchun No.4. In Jinyou No.1, CsPIP1:1 gene expression gradually increased to 2.3-fold at 4 h, followed by a decline in expression. The results indicated that ALA might delay and counteract the upregulated expression of CsPIP1:1 and CsNIP genes in cucumber seedlings under NaCl stress. Thus, salt tolerance of cucumber seedlings might be enhanced by ALA application.
Assuntos
Aquaporinas/biossíntese , Cucumis sativus/genética , Proteínas de Plantas/biossíntese , Estresse Fisiológico/genética , Ácido Aminolevulínico/farmacologia , Cucumis sativus/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Salinidade , Tolerância ao Sal/genética , Plântula/genética , ÁguaRESUMO
Adipose tissue plays an important role in energy metabolism and related diseases. The content of intramuscular fat significantly influences the pork quality. In this study, the whole gene expression of dorsal subcutaneous (s.c.) adipose tissue and intramuscular (i.m.) adipose tissue isolated from longissimus dorsi muscle tissue were compared using Affymetrix Gene-Chip microarray technology. The result revealed that 1228 genes were more highly expressed in s.c. adipose tissue, whereas 965 genes were higher expressed in i.m. adipose tissue. We found that the s.c. adipose tissue had a stronger capacity of lipid metabolism and fatty acid metabolism compared with i.m. adipose tissue, and angiopoietin-like-4, neuronatin, neuron-derived orphan receptor-1 alfa, and chloride intracellular channel 5 may play important roles in the regulation of fat deposition between i.m. and s.c. adipose tissues.
Assuntos
Tecido Adiposo/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Animais , Biologia Computacional/métodos , Masculino , Músculo Esquelético , Reprodutibilidade dos Testes , Transdução de Sinais , Gordura Subcutânea/metabolismo , SuínosRESUMO
With-no-lysine (K) kinase-4 (WNK4) is a newly cloned kinase-encoding gene that plays a crucial role in the maintenance of electrolyte homeostasis. Mutations of WNK4 can cause pseudohypoaldosteronism type α, an autosomal dominant disease characterized by hyperkalemia, metabolic acidosis and hypertension. We explored the expression and regulatory mechanism of WNK4 in the human kidneys, which is a key regulator of blood pressure. Expression of WNK4 was determined by RT-PCR. Transcription initiation site and regulatory elements in the promoter region of WNK4 were systematically analyzed with a combined set of experimental and bioinformatic methods. Using 5'-RACE, we have determined the transcription initiation site. We identified a number of putative cis-acting elements by analysis of the promoter region with the TRANSFAC-TESS software; these were subsequently confirmed with an electrophoresis mobility shift assay. As confirmed by a CAT-ELISA reporter assay, the promoter region of WNK4 has a high level of transcriptional activity. Several hormones, in particular dexamethasone, can suppress the level of WNK4 mRNA. These results have shed light on the regulatory mechanism of WNK4 expression in kidneys, as well as the influence of various hormones on expression levels. This should prove useful for studies on the roles of WNK4 in the pathogenesis of hypertension.