过量表达星星草(Puccinellia tenuiflora)液泡型H~+-ATP酶c亚基基因提高转基因酵母的耐盐性

植物液泡型H+-ATP酶(V-ATPase)在次级转运与应对多种逆境胁迫中具有重要作用。本研究克隆并解析了能够生长在盐碱地上的野生星星草V-ATPase c亚基基因(Put VHA-c)的功能。本研究利用荧光蛋白(GFP)标记结合内涵体染色的方法研究了Put VHA-c蛋白在酵母细胞中的定位,并利用转基因酵母分析了Put VHA-c基因在盐胁迫中的作用。共定位实验显示Put VHA-c-GFP融合蛋白主要定位在酵母细胞的内涵体上。Northern杂交和实时定量PCR显示在星星草悬浮细胞中Put VHA-c基因的表达能够被盐胁迫所诱导。研究结果结合过表达Put VHA-c的转基因酵母具有更高的耐盐性的发现,说明Put VHA-c基因参与着盐胁迫的应答。     

Essential role of Fkbp6 in male fertility and homologous chromosome pairing in meiosis

Meiosis is a critical stage of gametogenesis in which alignment and synapsis of chromosomal pairs occur, allowing for the recombination of maternal and paternal genomes. Here we show that FK506 binding protein (Fkbp6) localizes to meiotic chromosome cores and regions of homologous chromosome synapsis. Targeted inactivation of Fkbp6 in mice results in aspermic males and the absence of normal pachytene spermatocytes. Moreover, we identified the deletion of Fkbp6 exon 8 as the causative mutation in spontaneously male sterile as/as mutant rats. Loss of Fkbp6 results in abnormal pairing and misalignments between homologous chromosomes, nonhomologous partner switches, and autosynapsis of X chromosome cores in meiotic spermatocytes. Fertility and meiosis are normal in Fkbp6 mutant females. Thus, Fkbp6 is a component of the synaptonemal complex essential for sex-specific fertility and for the fidelity of homologous chromosome pairing in meiosis.     

Phosphate Fertilizers for Sugarcane Used at Pre-Planting (Phosphorus Fertilizer Application)

Phosphate applications are still rare in sugarcane cultivation and can be done with phosphorus fertilizers of residual over. This study aims to evaluate the agronomic efficiency of phosphate fertilization before sugarcane planting and its effect on sugarcane yield. The study was carried out over three crop cycles at the Jalles Machado sugar-mill in Goianésia, GO, Brazil. The treatments consisted of five different phosphorus sources (soluble and insoluble) applied at a rate of 300 kg ha^?1 phosphorus pentoxide (P₂O₅). The phosphorus fertilizers used were: triple superphosphate, mono-ammonium phosphate (MAP), Arad rock phosphate, Itafós rock phosphate, and magnesium term phosphate. The effects of these fertilizers were evaluated by plant height (ground to Top Visible Dewlap; TVD), leaf phosphorus content, agronomic efficiency, and sugarcane yield from cane-plants, first ratoon, and second ratoon. Magnesium term phosphate and triple superphosphate resulted in greater plant-cane height. Leaf phosphorus concentrations were highest with applications of MAP, magnesium term phosphate, and triple superphosphate. Phosphate applications did not affect sugarcane plant yield, but did increase ratoon yield. In general, the residual effect of insoluble phosphate increased over the years. The agronomic efficiency (AE) of phosphate was (in descending order): triple superphosphate (100%), term phosphate (89%), MAP (80%), Itafós rock phosphate (67%), and Arad rock phosphate (60%).     

Alternative Potash Fertilizer Source for Millet Crop

Millet is a grass that responds to potassium fertilization and Alfa 01 and Alfa 02 are potential sources of this nutrient. Thus, the aim of this study was to evaluate the potential of as potassium sources by measuring accumulation of this nutrient in millet. The crop was grown in Ustoxic Quartzipsamment (RQo) soil. The experiment was set up in randomized blocks with three repetitions. The treatments consisting of three sources (KCl, Alfa 02, Alfa 01), two rates (200 and 400 kg ha^?1 K₂O) and with no K fertilization (control). Potassium chloride was used as the standard to which the other sources of potassium were compared. Two consecutive crops of millet were grown in the same pots. The first crop received potassium source application but not the second. At the end of experiment, shoot dry matter production, shoot potassium concentration, shoot accumulated potassium, soil potassium and relative agronomic efficiency were determined. The relative agronomic efficiency, calculated from crops, was 100% for KCl, was 84% for Alfa 02 and was 11% for Alfa 01. In the shoot dry matter production and accumulated potassium only Alfa 01 was not statistically different of control. Millet absorbed potassium from the Alfa 02 source in both the first and second crops.     

Detection of Mycoplasma hyopneumoniae in lung and nasal swab samples from pigs by nested PCR and culture methods

We examined nasal swab and lung homogenate samples collected from pigs experimentally and naturally infected with Mycoplasma hyopneumoniae for the detection of M. hyopneumoniae by the nested PCR (nPCR) and culture methods. In the 23 experimentally infected pigs, M. hyopneumoniae was commonly detected in nasal swabs by the nPCR and culture methods at 4 weeks after inoculation, and there was a significant correlation (P<0.01) between the titers of viable organisms in nasal swabs and in lung homogenates in the experimentally inoculated pigs. In the naturally infected pigs, on the other hand, discrepancies in detection were found between nasal swab and lung homogenate samples in 17 of 36 cases, although the presence of gross lung lesions correlated relatively well with the detection of organisms from the samples. Our results indicated that the diagnosis of mycoplasmal pneumonia by nPCR in individual pigs with nasal swabs is reliable under these experimental conditions. At present, nPCR with nasal swabs should only be used for monitoring the disease status at the herd level under field conditions.     

Histological studies on the ontogeny of bovine palatine and pharyngeal tonsil: germinal center formation, IgG, and IgA mRNA expression

The development and distribution of lymphocyte subsets in calf palatine and pharyngeal tonsil were examined. During prenatal development, B cells were distributed in the subepithelial area, and T cells and MHC class II⁺ cells were found in the deep layer of B-cell area, respectively, in both tonsils. At neonatal stage, lymphoid follicle containing a few CD4⁺ cells have been formed in both tonsils. IgG⁺ and IgA⁺ cells were found in the parafollicular and epithelial area. At 3 months old, many germinal centers were recognized in both tonsils. CD4⁺ cells and IgG mRNA expression were detected in light zone of germinal centers. Many IgG, and IgA mRNA expressions also could be detected in the parafollicular and subepithelial area of both tonsils. The data suggest that both tonsils have an important role of local immune defense against invading antigen after birth. The comparison of the histological characteristics of tonsil and Peyer's patch during ontogeny is also discussed.     

Variable number of tandem aminoacid repeats in adhesion-related CDS products in Mycoplasma hyopneumoniae strains

The Mycoplasma hyopneumoniae genome contains at least 22 regions with a variable number of tandem nucleotide repeats (VNTRs) within coding DNA sequences (CDSs). In this work, the VNTR-containing CDSs were analysed in order to evaluate their degree of variation, possible correlations with antigenic properties, and their potential to be used as a basis for a strain typing PCR assay. We have analysed the VNTRs in five M. hyopneumoniae strains (J, 7448, 7422, PMS, and 232), based on published genomic sequences and on amplified and sequenced DNA segments. These VNTRs are distributed among 12 genes, most of which encode putative surface proteins, including known adhesins. The number of repeat units in any of the VNTRs is highly variable among the analysed strains, but they are, without exception, translationally in frame, and, therefore, code for a variable number of aminoacid repeats (VNTARs). These VNTARs determine putative structural, physicochemical and antigenic variations in the corresponding proteins, with potential implications for aspects associated to M. hyopneumoniae pathogenicity, such as cell adhesion and interactions with the host immune system. Considering that the characterized VNTARs are relatively stable, at least in vitro, and their sizes are strain-specific, we have developed a VNTR-based PCR assay for M. hyopneumoniae strain identification, useful for enzootic pneumonia (EP) diagnosis, strain typing, and distinction of circulating field isolates from vaccine strains in animals vaccinated against EP.