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1.
Genetic relationships among 18 accessions, including 16 of Ananas and two of Pseudananas, were investigated using RAPD molecular markers. The procedure for DNA extraction was adapted from the method of Dellaporta et al. (1983) where an incubation in proteinase K and a purification step were included. From the total of 148 markers scored,132 (89.2%) were polymorphic. The similarity matrix was used for cluster analysis. The phenogram developed from the RAPD bands showed that for most of the cases, the accessions within a species grouped together. Nevertheless, a moderate infraspecific genetic variation was observed. For example, DNA data grouped all A. comosus accessions with a mean similarity coefficient of 0.85. Comparable results were obtained with all other species investigated. The highest genetic divergence was found withinA. lucidus where the mean similarity coefficient among accessions was0.75. A similar level of genetic polymorphism was observed among species,therefore, a definition about which species were involved in the constitution of A. comosus genotypes was not possible. These results agree with the breeders standpoint suggesting that all Ananas species belong to the primary gene pool of pineapple. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   
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Purification of chicken anemia virus (CAV) VP3 protein, expressed in a prokaryotic expression system as histidine-tagged fusion protein is demonstrated in the present study. CAV particle was obtained from infected liver of chicken and DNA was extracted. The VP3 protein gene was amplified from the extracted DNA by polymerase chain reaction (PCR) and cloned. The recombinant expression construct (pTrc-VP3) was identified by PCR and sequencing analysis. Expression of VP3 protein with a molecular mass of approximately 21kDa was confirmed by Western blotting analysis with CAV-specific antibodies. The in vitro expressed VP3 protein was purified to near homogeneity by elution from the gel, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The purified VP3 protein was recognized by CAV antibodies in a Western blotting assay. This finding indicates that recombinant VP3 expressed in the pTrcHis2 vector system can be used as antigen to detect anti-CAV antibodies.  相似文献   
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The petroleum industry activities provide potential risks to the environment because they can contaminate ecosystems with different organic compounds in the production chain. Several accidents with transport and handling of petroleum and related products occurred in urban areas with harmful effects to the quality of life and economy. In the 1990s, bioremediation and phytoremediation technologies as economically feasible alternatives to repair the environmental damage were developed. In this study, the potential of the willows Salix rubens and Salix triandra were evaluated with regard to the phytoremediation of soils contaminated with petroleum-derived hydrocarbons (total hydrocarbons and polycyclic aromatic hydrocarbons (PAHs)). The PAHs were quantified by extraction from soils and plants using dichloromethane under ultrasonication. The HPLC analysis was performed with GC/MSD equipment. The total hydrocarbons present in uncontaminated soil were quantified by the sum of animal/vegetable oils and greases and mineral oils and greases according to Standard Methods 5520 (1997). The two willows species S. rubens and S. triandra were resistant during the project development. In the contaminated soil, in which both species were planted, the total hydrocarbons concentration was reduced near 98?%. The PAHs content was remarkably reduced as well. Pyrene showed an initial concentration of 23.06???g?kg?1, decreasing in most cases to 0.1???g?kg?1 or to undetectable levels. Chrysene decreased from 126.27???g?kg?1 to undetectable levels. Benzo[k]fluoranthene and benzo[a]pyrene concentrations had also showed a decrease from 28.44 and 3.82???g?kg?1, respectively, to undetectable levels.  相似文献   
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1. Infectious laryngotracheitis is a respiratory disease that affects the poultry industry worldwide. It is common in flocks with high-bird density, causing major economic losses.

2. In this study, a SYBR® FAST polymerase chain reaction (PCR) double-strand DNA intercalating agent assay was performed for the detection of infectious laryngotracheitis virus (ILTV) in clinical samples in comparison with a conventional nested-PCR, both based on the glycoprotein E encoding gene. This assay amplified 56 bp and was capable of detecting 19 to 1 copies of virus.

3. In total, 164 clinical samples were obtained from birds with respiratory problems from the period of 2009–2016. In the nested-PCR, there were 45.12% positive samples and 54.88% negative samples, while in the real-time PCR (qPCR), there were 81.1% positive samples and 18.9% negative samples.

4. In conclusion, qPCR from the DNA double-strand intercalating agent SYBR® GREEN FAST was useful for the diagnosis of ILTV because it detected samples that were negative in nested-PCR. This assay has advantages, such as a shortened processing-time, and no need for post-amplification processing (electrophoresis) with additional reagents, such as MgCl2 and agarose. Hence, qPCR proved to be useful, rapid and low cost for use with clinical samples.  相似文献   

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Tropical Animal Health and Production - The objective of this study was to evaluate the effect of feeding Saanen goat kids with calcium salts of fatty acids (CSFA) in diet, on intake, performance,...  相似文献   
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Recent reports of weed‐control failures after the use of glyphosate led to suspicion about the selection of resistant biotypes of Conyza at locations in west and north Paraná, Brazil. Plants were collected, identified as Conyza sumatrensis and subsequently evaluated for possible resistance to glyphosate in four stages of weed development. The experiments were carried out in a greenhouse by combining biotypes, stages of development and a range of glyphosate doses. All the suspected biotypes were collected from locations in Cascavel, Toledo, Assis Chateaubriand, Tupãssi and Campo Mourão with a history of glyphosate use in burndown and in glyphosate‐resistant soybean for at least the four previous years and were compared to a susceptible biotype (São Jorge do Ivaí) with no previous history of herbicide use. The doses of glyphosate ranged from 0 to 5760 g ae ha?1. The biotypes were considered as resistant if two combined criteria were present (resistance factor > 1 and the rate required to achieve 80% control is >720 g ha?1). The results provided evidence that there is a marked difference in the level of control of older plants and also confirmed the presence of some resistant biotypes. For applications at the first stage of development, two biotypes that were resistant to glyphosate were identified (Cascavel‐1 and Tupãssi‐6). For applications in the second stage of development, beyond the biotypes that were found in the first stage, three other biotypes were considered as resistant: Toledo‐5, Assis Chateaubriand‐7 and Floresta‐10. However, for applications at the third and fourth stages, all the biotypes were considered as resistant.  相似文献   
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1. Infectious bronchitis virus (IBV) variants in Brazil were isolated during 2010–2015 for epidemiological and molecular analysis to characterise the different variants and perform a bioinformatic analysis to compare with sequences of variants collected over the previous 40 years.

2. Of the 453 samples examined, 61.4% were positive for IBV and 75.9% of these were considered to have the BR-I genotype and were detected in birds of all ages distributed in all five Brazilian regions.

3. The ratio of non-synonymous substitutions per non-synonymous site (dN) to synonymous substitutions per synonymous site (dS), i.e. dN/dS, revealed a predominance of codons with non-synonymous substitutions in the first third of the S1 gene and a dN/dS ratio of 0.67. Additionally, prediction of N-glycosylation sites showed that most of the BR-I variants (from 2003 to early 2014) had an extra site at amino acid position 20, whereas the newest variants lacked this extra site.

4. These results suggest that Brazilian IBV variants probably underwent drastic mutations at various points between 1983 and 2003 and that the selection processes became silent after achieving a sufficiently effective antigenic structure for invasion and replication in their hosts. Brazilian IBV genotype BR-I is currently the predominant genotype circulating in Brazil and South America.  相似文献   

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The objective of the present work was to develop species-specific microsatellite markers for P. scalare and to analyze the diversity and genetic structure of a wild population, from the Amazon River, and three commercial stocks (common, marble, and clown morphological variants), from farmers in Vieras-Minas Gerais. Through microsatellite-enriched genetic libraries, 11 microsatellite markers with adequate amplification patterns were characterized. Population genetic analysis identified eight polymorphic loci that generated 66 alleles ranging from two alleles (PSCA1B3) to nine (PSCA2H1). The polymorphic information content ranged from 0.031 to 0.827. High genetic differentiation was observed between the wild population and the stocks, and moderate differentiation between the three stocks. Deviation in the Hardy-Weinberg equilibrium was observed in one locus in the wild population, in five loci in the common morphological variant, in two in the marble, and in two in the clown morphological variant. Bayesian analysis of genetic structure revealed the existence of two clusters, one represented by the natural population and the other by the stocks. The developed microsatellite markers serve as a tool for the analysis of diversity and genetic structure and conservation studies of P. scalare.

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