RESUMO
Protein ubiquitination is extensively involved in the regulation of a considerable number of physiological processes in plant cells. E2 (ubiquitin-conjugating enzyme, UBC), one of the essential enzymes of eukaryotic ubiquitination, catalyzes protein ubiquitination together with E1 and E3. In this study, we cloned four full-length cDNA NnUBCs of Nelumbo nucifera. With the same coding sequence length of 459 bp and coding 153 amino acids, these four genes are highly homologous with the AtUBC1 and AtUBC2 of Arabidopsis thaliana. Quantitative fluorescence polymerase chain reaction showed that these four genes exhibited different expression patterns in different tissues of N. nucifera. Overall, the expression of NnUBC3 was the highest in all plant tissues. Tests of different stress treatments showed that NnUBC3 plays an important role in response to heat, salt, and drought stresses in N. nucifera. Moreover, transgenic Arabidopsis plants (Atubc1-1Atubc2-1 mutant) expressing NnUBC3 presented a wild-type phenotype, indicating that NnUBC3 performs the same function as AtUBC1 and AtUBC2.
Assuntos
Nelumbo/enzimologia , Proteínas de Plantas/genética , Enzimas de Conjugação de Ubiquitina/genética , Sequência de Aminoácidos , Arabidopsis , Sequência de Bases , Clonagem Molecular , Expressão Gênica , Nelumbo/genética , Filogenia , Proteínas de Plantas/metabolismo , Homologia de Sequência de Aminoácidos , Estresse Fisiológico , Enzimas de Conjugação de Ubiquitina/metabolismoRESUMO
Water chestnut (Eleocharis dulcis) is an important aquatic crop in China; however, transcriptomic and genomic data in public databases are limited. To identify genes and development molecular markers, high-throughput transcriptome sequencing was applied to generate transcript sequences from water chestnut leaf. More than 24 million reads were obtained, trimmed, and assembled into 40,796 contigs with an average length of 616.6 bp. Sequence similarity analyses against 4 public databases (NR, GO, KEGG, KOG) revealed 17,628 contigs that could be annotated with gene descriptions, conserved protein domains, or gene ontology terms. Among the important metabolic pathways, 27 genes were related to starch synthesis and 13 genes were in the steroid synthetic pathway. In addition, 2570 cDNA simple sequence repeats were identified as potential molecular markers in our contigs. One hundred pairs of polymerase chain reaction primers were designed and used for validation of the amplification. The results revealed that 87 primer pairs were successfully amplified in initial screening tests. Overall, this transcriptome dataset and these markers can serve as a platform for further gene expression studies, functional genomic studies, and marker-assisted selection in E. dulcis.
Assuntos
Eleocharis/genética , Repetições de Microssatélites/genética , Folhas de Planta/genética , Transcriptoma/genética , China , DNA Complementar/genética , Eleocharis/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Sequenciamento de Nucleotídeos em Larga Escala , Anotação de Sequência MolecularRESUMO
Nelumbo nucifera is an important economic vegetable and traditional medicine, but available genetic resources remain limited. Next generation sequencing has proven to be a rapid and effective means of identifying genic simple sequence repeat (genic-SSR) markers. This study developed genic-SSRs for N. nucifera using Illumina sequencing technology to assess diversity across cultivated and wild lotus. A total of 105,834 uni-contigs were produced with an average read length of 722 bp. Exactly 11,178 genic-SSR loci were identified in 9523 uni-contigs. Di-nucleotide (64.5%) was the most abundant SSR, followed by tri-nucleotide (23%), tetra-nucleotide (8.9%), penta-nucleotide (2.5%), and hexa-nucleotide (1%) repeat types. The most common di- and tri-nucleotide repeat motifs were AG/CT (51%) and AAG/CTT (8%), respectively. Based on these SSRs sequences, 6568 primer pairs were designed, of which 72 primers were randomly selected for synthesis and validation, and 38 in-silico polymorphic primers were obtained using in-house perl scripts. A total of 110 primers were screened in the lotus samples and the results showed that 101 primers yielded amplification products, of which 80 were polymorphs. The number of alleles ranged from 2 to 17 and the PIC (polymorphism information content) ranged from 0.19 to 0.87 with a mean value of 0.55. An Unweighted Pair Group Method with Arithmetic Mean (UPGMA) dendrogram based on Jaccard's similarity coefficients showed that the correlation between geographical source and genotype was low. This study describes the distribution of genic-SSRs in the expressed portion of the lotus genome. These genic-SSRs have an important role to play in molecular mapping, diversity analysis, and marker-assisted selection strategies in Nelumbo.
Assuntos
Genoma de Planta , Nelumbo/genética , RNA de Plantas/genética , Flores/genética , Frequência do Gene , Loci Gênicos , Marcadores Genéticos , Repetições de Microssatélites , Filogenia , Polimorfismo Genético , Rizoma/genética , Análise de Sequência de RNA , Especificidade da EspécieRESUMO
Taurine (2-aminoethanesulfonic acid) is widely distributed in animal tissues and has diverse pharmacological effects. However, the role of taurine in modulating smooth muscle contractility is still controversial. We propose that taurine (5-80 mM) can exert bidirectional modulation on the contractility of isolated rat jejunal segments. Different low and high contractile states were induced in isolated jejunal segments of rats to observe the effects of taurine and the associated mechanisms. Taurine induced stimulatory effects on the contractility of isolated rat jejunal segments at 3 different low contractile states, and inhibitory effects at 3 different high contractile states. Bidirectional modulation was not observed in the presence of verapamil or tetrodotoxin, suggesting that taurine-induced bidirectional modulation is Ca2+ dependent and requires the presence of the enteric nervous system. The stimulatory effects of taurine on the contractility of isolated jejunal segments was blocked by atropine but not by diphenhydramine or by cimetidine, suggesting that muscarinic-linked activation was involved in the stimulatory effects when isolated jejunal segments were in a low contractile state. The inhibitory effects of taurine on the contractility of isolated jejunal segments were blocked by propranolol and L-NG-nitroarginine but not by phentolamine, suggesting that adrenergic β receptors and a nitric oxide relaxing mechanism were involved when isolated jejunal segments were in high contractile states. No bidirectional effects of taurine on myosin phosphorylation were observed. The contractile states of jejunal segments determine taurine-induced stimulatory or inhibitory effects, which are associated with muscarinic receptors and adrenergic β receptors, and a nitric oxide associated relaxing mechanism.
Assuntos
Animais , Masculino , Jejuno/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Miosinas/metabolismo , Taurina/farmacologia , Antagonistas Adrenérgicos alfa/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Atropina/farmacologia , Bloqueadores dos Canais de Cálcio/farmacologia , Cimetidina/farmacologia , Difenidramina/farmacologia , Sistema Nervoso Entérico/efeitos dos fármacos , Antagonistas dos Receptores Histamínicos H1/farmacologia , /farmacologia , Jejuno/fisiologia , Antagonistas Muscarínicos/farmacologia , Quinase de Cadeia Leve de Miosina/metabolismo , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico/metabolismo , Fosforilação , Fentolamina/farmacologia , Propranolol/farmacologia , Ratos Sprague-Dawley , Taurina/antagonistas & inibidores , Tetrodotoxina/farmacologia , Verapamil/farmacologiaRESUMO
Taurine (2-aminoethanesulfonic acid) is widely distributed in animal tissues and has diverse pharmacological effects. However, the role of taurine in modulating smooth muscle contractility is still controversial. We propose that taurine (5-80 mM) can exert bidirectional modulation on the contractility of isolated rat jejunal segments. Different low and high contractile states were induced in isolated jejunal segments of rats to observe the effects of taurine and the associated mechanisms. Taurine induced stimulatory effects on the contractility of isolated rat jejunal segments at 3 different low contractile states, and inhibitory effects at 3 different high contractile states. Bidirectional modulation was not observed in the presence of verapamil or tetrodotoxin, suggesting that taurine-induced bidirectional modulation is Ca(2+) dependent and requires the presence of the enteric nervous system. The stimulatory effects of taurine on the contractility of isolated jejunal segments was blocked by atropine but not by diphenhydramine or by cimetidine, suggesting that muscarinic-linked activation was involved in the stimulatory effects when isolated jejunal segments were in a low contractile state. The inhibitory effects of taurine on the contractility of isolated jejunal segments were blocked by propranolol and L-NG-nitroarginine but not by phentolamine, suggesting that adrenergic ß receptors and a nitric oxide relaxing mechanism were involved when isolated jejunal segments were in high contractile states. No bidirectional effects of taurine on myosin phosphorylation were observed. The contractile states of jejunal segments determine taurine-induced stimulatory or inhibitory effects, which are associated with muscarinic receptors and adrenergic ß receptors, and a nitric oxide associated relaxing mechanism.
Assuntos
Jejuno/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Miosinas/metabolismo , Taurina/farmacologia , Antagonistas Adrenérgicos alfa/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Animais , Atropina/farmacologia , Bloqueadores dos Canais de Cálcio/farmacologia , Cimetidina/farmacologia , Difenidramina/farmacologia , Sistema Nervoso Entérico/efeitos dos fármacos , Antagonistas dos Receptores Histamínicos H1/farmacologia , Antagonistas dos Receptores H2 da Histamina/farmacologia , Jejuno/fisiologia , Masculino , Antagonistas Muscarínicos/farmacologia , Quinase de Cadeia Leve de Miosina/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/antagonistas & inibidores , Fentolamina/farmacologia , Fosforilação , Propranolol/farmacologia , Ratos Sprague-Dawley , Taurina/antagonistas & inibidores , Tetrodotoxina/farmacologia , Verapamil/farmacologiaRESUMO
The protective effects of remifentanil preconditioning on serum superoxide dismutase (SOD) and malondialdehyde (MDA) during pump-assisted coronary artery bypass graft (CABG) were investigated. Forty pump-assisted CABG patients were randomly divided into a remifentanil preconditioning group (R group) and a control group (C group, N = 10; normal saline). The R group was further divided into 3 sub-groups (R1, R2, and R3; N = 10 per group) according to the remifentanil dose (0.6, 1.2, and 1.8 µg·kg(-1)·min(-1), respectively). A venous blood sample was taken at anesthesia induction (T0), before cardiopulmonary bypass (CPB) (T1), CPB 30 min (T2), and after CPB (T3), and protein concentrations were measured. Patients were tested 24 h before and after the operation with the Mini-Mental State Examination (MMSE), and the difference was calculated. The MMSE score difference in the R3 group was lower than those of the other 3 groups (P < 0.05). At T2 and T3, the R3 group showed a significant decrease in S-100ß protein and MDA and an increase in SOD (P < 0.05) compared with the other groups, and S-100ß was negatively correlated with SOD activity (T2: r = -0.76, -0.80, P < 0.01; T3: r = -0.795, P < 0.01), and was positively correlated with MDA density (T2: r = 0.71, P < 0.01; T3: r = 0.71, P < 0.01). In conclusion, high-dosage remifentanil preconditioning played a protective role on brain damage, possibly through inhibition of the oxidative stress response.
Assuntos
Lesões Encefálicas/prevenção & controle , Ponte de Artéria Coronária/métodos , Hipnóticos e Sedativos/administração & dosagem , Piperidinas/administração & dosagem , Pré-Medicação , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , RemifentanilRESUMO
Amorphophallus konjac is an herbaceous tuber crop with tremendous potential for commercial development. We report the development of microsatellite primers for this important crop species. Thirteen polymorphic microsatellite markers were developed and tested in two populations of A. konjac from the Wuling Mountain Region (WL population) and the Yunnan Province (YN population) in China. The number of alleles per locus ranged from 1 to 7; the observed and expected heterozygosities ranged from 0 to 1 and from 0 to 0.844, respectively, in the two populations. These microsatellite markers will facilitate further studies in population genetics and utilization of A. konjac.