RESUMO
Molecular cloning of thyrotropin-releasing hormone receptors (TRHR) was performed in a teleost, the sockeye salmon (Oncorhynchus nerka). Four different TRHR cDNAs were cloned and named TRHR1, TRHR2a, TRHR2b and TRHR3 based on their similarity to known TRHR subtypes in vertebrates. Important residues for TRH binding were conserved in deduced amino acid sequences of the three TRHR subtypes except for the TRHR2b. Seven transmembrane domains were predicted for TRHR1, TRHR2a and TRHR3 proteins but only five for TRHR2b which appears to be truncated. In silico database analysis identified putative TRHR sequences including invertebrate TRHR and reptilian, avian and mammalian TRHR3. Phylogenetic analyses predicted the molecular evolution of TRHR in vertebrates: from the common ancestral TRHR (i.e. invertebrate TRHR), the TRHR2 subtype diverged first and then TRHR1 and TRHR3 diverged. Reverse transcription-polymerase chain reaction analyses revealed TRHR1 transcripts in the brain (hypothalamus), retina, pituitary gland and large intestine; TRHR2a in the brain (telencephalon and hypothalamus); and TRHR3 in the brain (olfactory bulbs) and retina.
Assuntos
DNA Complementar/genética , Proteínas de Peixes/genética , Receptores do Hormônio Liberador da Tireotropina/genética , Salmão/genética , Sequência de Aminoácidos , Animais , Clonagem Molecular , Evolução Molecular , Proteínas de Peixes/química , Proteínas de Peixes/metabolismo , Expressão Gênica/genética , Dados de Sequência Molecular , Receptores do Hormônio Liberador da Tireotropina/química , Receptores do Hormônio Liberador da Tireotropina/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de ProteínaRESUMO
Recombinant Rhodopseudomonas palustris, harboring the carotenoid-metabolizing gene crtI (CrtIBS), and whose color changes from greenish yellow to red in response to inorganic As(III), was cultured in transparent microplate wells illuminated with a light emitting diode (LED) array. The cells were seen to grow better under near-infrared light, when compared with cells illuminated with blue or green LEDs. The absorbance ratio of 525 to 425 nm after cultivation for 24 h, which reflects red carotenoid accumulation, increased with an increase in As(III) concentrations. The detection limit of cultures illuminated with near-infrared LED was 5 microgram/l, which was equivalent to that of cultures in test tubes illuminated with an incandescent lamp. A near-infrared LED array, in combination with a microplate, enabled the simultaneous handling of multiple cultures, including CrtIBS and a control strain, for normalization by the illumination of those with equal photon flux densities. Thus, the introduction of a near-infrared LED array to the assay is advantageous for the monitoring of arsenic in natural water samples that may contain a number of unknown factors and, therefore, need normalization of the reporter event.
Assuntos
Arsênio/análise , Técnicas Biossensoriais/métodos , Monitoramento Ambiental/métodos , Rodopseudomonas/metabolismo , Poluentes Químicos da Água/análise , Arsênio/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Técnicas Biossensoriais/instrumentação , Carotenoides/metabolismo , Monitoramento Ambiental/instrumentação , Luz , Fotossíntese , Rodopseudomonas/genética , Poluentes Químicos da Água/metabolismoRESUMO
To assess the effects of tropospheric O3 on rice cultivated in Bangladesh, four Bangladeshi cultivars (BR11, BR14, BR28 and BR29) of rice (Oryza sativa L.) were exposed daily to charcoal-filtered air or O3 at 60 and 100 nl l(-1) (10:00-17:00) from 1 July to 28 November 2008. The whole-plant dry mass and grain yield per plant of the four cultivars were significantly reduced by the exposure to O3. The exposure to O3 significantly reduced net photosynthetic rate of the 12th and flag leaves of the four cultivars. The sensitivity to O3 of growth, yield and leaf gas exchange rates was not significantly different among the four cultivars. The present study suggests that the sensitivity to O3 of yield of the four Bangladeshi rice cultivars is greater than that of American rice cultivars and is similar to that of Japanese rice cultivars and that O3 may detrimentally affect rice production in Bangladesh.
Assuntos
Poluentes Atmosféricos/toxicidade , Oryza/efeitos dos fármacos , Ozônio/toxicidade , Folhas de Planta/efeitos dos fármacos , Transpiração Vegetal/efeitos dos fármacos , Poluentes Atmosféricos/metabolismo , Bangladesh , Oryza/crescimento & desenvolvimento , Oryza/metabolismo , Ozônio/metabolismo , Folhas de Planta/metabolismoRESUMO
The endogenous brassinosteroids in the dwarf mutant lk of pea (Pisum sativum) were quantified by gas chromatography-selected ion monitoring. The levels of castasterone, 6-deoxocastasterone, and 6-deoxotyphasterol in lk shoots were reduced 4-, 70-, and 6-fold, respectively, compared with those of the wild type. The fact that the application of brassinolide restored the growth of the mutant indicated that the dwarf mutant lk is brassinosteroid deficient. Gas chromatography-selected ion monitoring analysis of the endogenous sterols in lk shoots revealed that the levels of campestanol and sitostanol were reduced 160- and 10-fold, respectively, compared with those of wild-type plants. These data, along with metabolic studies, showed that the lk mutant has a defect in the conversion of campest-4-en-3-one to 5alpha-campestan-3-one, which is a key hydrogenation step in the synthesis of campestanol from campesterol. This defect is the same as that found in the Arabidopsis det2 mutant and the Ipomoea nil kbt mutant. The pea gene homologous to the DET2 gene, PsDET2, was cloned, and it was found that the lk mutation would result in a putative truncated PsDET2 protein. Thus it was concluded that the short stature of the lk mutant is due to a defect in the steroidal 5alpha-reductase gene. This defect was also observed in the callus induced from the lk mutant. Biosynthetic pathways involved in the conversion of campesterol to campestanol are discussed in detail.
Assuntos
3-Oxo-5-alfa-Esteroide 4-Desidrogenase/genética , Colesterol/análogos & derivados , Pisum sativum/genética , Sequência de Aminoácidos , Brassinosteroides , Colestanóis/metabolismo , Colesterol/metabolismo , Sequência Conservada , Nanismo/genética , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Mutação/genética , Pisum sativum/enzimologia , Fitosteróis/metabolismo , Doenças das Plantas , Alinhamento de Sequência , Deleção de Sequência , Homologia de Sequência de Aminoácidos , Sitosteroides/metabolismo , Esteroides Heterocíclicos/metabolismoRESUMO
Cadmium pollution in soil not only affects growth and yield formation of crop plants, but also develops a potential risk to food health. The experiment was conducted to study growth of Cd and mineral nutrient uptake of two wheat cultivars differed in agronomic performance in seedling stage under 5 Cd levels ranged from 0 to 1.0 mg.kg-1 Cd. The results showed that the growth and dry matter accumulation were stimulated in low Cd concentration (0.03 mg.kg-1) for both cultivars, while inhibited significantly under high concentration (> 0.3 mg.kg-1). The inhibited extension was dependence on cultivars and Genguo 534 had higher tolerance to Cd toxicity in comparison with E18513. There was the significant interaction between Cd level and cultivar in the influence of Cd treatment on Cd content in both root and shoot in that Genguo 534 had higher Cd content in lower Cd levels and E81513 showed more rapid increase in higher Cd levels. Cd treatment had significant influence on mineral nutrient uptake and two cultivars showed different response. In the treatment of 0.1 mg.kg-1 Cd, the contents of five macro-nutrients were significant lower for E81513 in comparison with those in control, while Genguo remained little changed.