Faba beans are inexpensive, nutrient-dense sources of plant protein, but anti-nutritional factors such as condensed tannins
reduce the biological value of their protein. Two recessive genes, zt-1 and zt-2, control the absence of tannins in faba bean seeds and also determine a white flower character on the plant. However, crosses
between them produce coloured F1 plants with tannins that contaminate the crop. Therefore, it is important to identify the gene present in all tannin-free
cultivars and gene bank accessions to enable breeders to choose appropriate genitors for their crosses. The aim of this study
was the identification of markers linked to zt-1, one of the genes governing free tannin content in faba bean. A segregating F2 population derived from the cross between the coloured flower and high tannin content genotype Vf6 and a zt-1 line was developed and characterized phenotypically. Bulked Segregant Analysis (BSA) was used to identify Random Amplified
Polymorphic DNA (RAPD) markers linked to the zt-1 gene. Four RAPD loci (OPC5551, OPG15600, OPG111171 and OPAF20776) showed polymorphism between the contrasting bulks. The markers were sequenced to develop specific Sequence Characterised
Amplified Regions (SCARs). Amplification of SCC5551 produced a single product which was only observed in the white flowered and zero tannin content genotypes, whereas SCAR
SCG111171only produced a band in F2 plants with coloured flower and high tannin content. SCARs SCC5551 and SCG111171 were tested for their applicability for routine screening in 37 faba bean genotypes differing in flower colour and tannin
content. SCC5551, allowed the prediction of the zt-1 genotypes with a 95% of accuracy, underscoring the potential of this SCAR marker as a cost-effective tool for MAS in large
faba bean breeding populations. 相似文献
The axonal microtubule‐associated protein tau is a well‐known regulator of microtubule stability in neurons. However, the putative interplay between tau and End‐binding proteins 1 and 3 (EB1/3), the core microtubule plus‐end tracking proteins, has not been elucidated yet. Here, we show that a cross‐talk between tau and EB1/3 exists in developing neuronal cells. Tau and EBs partially colocalize at extending neurites of N1E‐115 neuroblastoma cells and axons of primary hippocampal neurons, as shown by confocal immunofluorescence analyses. Tau down‐regulation leads to a reduction of EB1/3 comet length, as observed in shRNA‐stably depleted neuroblastoma cells and TAU?/? neurons. EB1/3 localization depends on the expression levels and localization of tau protein. Over‐expression of tau at high levels induces EBs relocalization to microtubule bundles at extending neurites of N1E‐115 cells. In differentiating primary neurons, tau is required for the proper accumulation of EBs at stretches of microtubule bundles at the medial and distal regions of the axon. Tau interacts with EB proteins, as shown by immunoprecipitation in different non‐neuronal and neuronal cells and in whole brain lysates. A tau/EB1 direct interaction was corroborated by in vitro pull‐down assays. Fluorescence recovery after photobleaching assays performed in neuroblastoma cells confirmed that tau modulates EB3 cellular mobility. In summary, we provide evidence of a new function of tau as a direct regulator of EB proteins in developing neuronal cells. This cross‐talk between a classical microtubule‐associated protein and a core microtubule plus‐end tracking protein may contribute to the fine‐tuned regulation of microtubule dynamics and stability during neuronal differentiation.
Rodent ovariectomy is an experimental method to eliminate the main source of sexual
steroids. This work explored for the first time the ovariectomy temporal changes induced
in the hemostatic coagulation markers: prothrombin time (PT), activated partial
thromboplastin time (aPTT), thrombin time (TT), and fibrinogen concentration (FIB) along
with uterine weight on adult female CD1 mice and Wistar rats. Uterine weight (Uw) was
assessed before ovariectomy (control), and 1, 3, 5, 7, 9, 16, and 21 days after surgery.
PT, aPTT, TT and FIB were estimated the same days, using reported standard techniques.
Ovariectomy decreased Uw, since day 1; and from day 10 to 21 reached the lowest values for
both species. After day 1, mice hemostatic parameters changed (PT +10%,
P<0.05; aPTT +53%, P<0.05; TT −24%,
P<0.05; FIB +67%, P<0.05). Rats showed
significant changes only in TT and FIB (TT −13%, P<0.001; FIB +65%,
P<0.001). Neither mice PT, aPTT and TT, recovered control values
after 21 days. In the rats from day 5 to 16 aPTT diminished (18–23%,
P<0.05) recovering to control values on day 21, TT after 9 days and PT
on day 16. In both species, FIB returned to its control values after 9 days. Ovariectomy
differentially altered the PT hemostatic parameter of mice and rats indicating a
non-equivalence among both species behaviour for experimental studies of blood
coagulation. 相似文献
The functional role of burst firing (i.e. the firing of packets of action potentials followed by quiescence) in sensory processing is still under debate. Should bursts be considered as unitary events that signal the presence of a particular feature in the sensory environment or is information about stimulus attributes contained within their temporal structure? We compared the coding of stimulus attributes by bursts in vivo and in vitro of electrosensory pyramidal neurons in weakly electric fish by computing correlations between burst and stimulus attributes. Our results show that, while these correlations were strong in magnitude and significant in vitro, they were actually much weaker in magnitude if at all significant in vivo. We used a mathematical model of pyramidal neuron activity in vivo and showed that such a model could reproduce the correlations seen in vitro, thereby suggesting that differences in burst coding were not due to differences in bursting seen in vivo and in vitro. We next tested whether variability in the baseline (i.e. without stimulation) activity of ELL pyramidal neurons could account for these differences. To do so, we injected noise into our model whose intensity was calibrated to mimic baseline activity variability as quantified by the coefficient of variation. We found that this noise caused significant decreases in the magnitude of correlations between burst and stimulus attributes and could account for differences between in vitro and in vivo conditions. We then tested this prediction experimentally by directly injecting noise in vitro through the recording electrode. Our results show that this caused a lowering in magnitude of the correlations between burst and stimulus attributes in vitro and gave rise to values that were quantitatively similar to those seen under in vivo conditions. While it is expected that noise in the form of baseline activity variability will lower correlations between burst and stimulus attributes, our results show that such variability can account for differences seen in vivo. Thus, the high variability seen under in vivo conditions has profound consequences on the coding of information by bursts in ELL pyramidal neurons. In particular, our results support the viewpoint that bursts serve as a detector of particular stimulus features but do not carry detailed information about such features in their structure. 相似文献
Engineering proteins for selective tissue targeting can improve therapeutic efficacy and reduce undesired side effects. The relatively high dose of recombinant human acid α-glucosidase (rhGAA) required for enzyme replacement therapy of Pompe disease may be attributed to less than optimal muscle uptake via the cation-independent mannose 6-phosphate receptor (CI-MPR). To improve muscle targeting, Zhu et al. (1) conjugated periodate oxidized rhGAA with bis mannose 6-phosphate bearing synthetic glycans and achieved 5-fold greater potency in a murine Pompe efficacy model. In the current study, we systematically evaluated multiple strategies for conjugation based on a structural homology model of GAA. Glycan derivatives containing succinimide, hydrazide, and aminooxy linkers targeting free cysteine, lysines, and N-linked glycosylation sites on rhGAA were prepared and evaluated in vitro and in vivo. A novel conjugation method using enzymatic oxidation was developed to eliminate side oxidation of methionine. Conjugates derived from periodate oxidized rhGAA still displayed the greatest potency in the murine Pompe model. The efficiency of conjugation and its effect on catalytic activity were consistent with predictions based on the structural model and supported its use in guiding selection of appropriate chemistries. 相似文献
The Liolaemus lineomaculatus section is a geographically widely distributed group of lizards from the Patagonian region of southern South America, and includes 18 described species representing the most southerly distributed Liolaemus taxa (the genus includes 228 species and extends from Tierra del Fuego north to south-central Peru). Despite high species diversity, the phylogenetic relationships of this section are unknown. In the present work we sampled all described species in the L. lineomaculatus section as well as currently undescribed candidate species to reconstruct the first complete phylogenetic hypothesis for the clade. Our data set included four anonymous nuclear loci, three nuclear protein-coding loci, and two mitochondrial genes. We compared results obtained with three different phylogenetic methods for the concatenated data set (Maximum Parsimony, Maximum Likelihood and Bayesian Inference) with a coalescent-based species tree approach (BEST), and recovered congruent, strongly-supported topological arrangements across all methods. We identified four main clades within the L. lineomaculatus section: the lineomaculatus, magellanicus, somuncurae, and kingii+archeforus groups, for which we estimated divergence times. We discuss the taxonomic implications of these results and how the future integration of phylogeographic, niche modeling and morphological approaches will allow testing biogeographical hypotheses in this clade. 相似文献
The lizard genus Liolaemus is endemic to temperate South America and includes more than 225 species. Liolaemus gracilis and L. bibronii are closely related species that have large and overlapping geographic distributions, and the objective of this work is to further investigate the L. bibronii–L. gracilis mtDNA paraphyletic pattern previously detected, using an integrative approach, based on mtDNA, nuclear DNA and morphological characters. We identified eight morphological L. bibronii introgressed with L. gracilis mtDNAs, and the reciprocal for one L. gracilis, from six localities in the region of sympatry overlap. The morphological identity of these introgressed individuals was confirmed by diagnostic nuclear markers, and this represents the first well-documented case of interspecific hybridization in the lizard genus Liolaemus. Of the three most likely hypotheses for these observed patterns, we suggest that asymmetrical mtDNA introgression as a result of recent or ongoing hybridization between L. bibronii and L. gracilis is the most likely. This may be due to size selection by L. gracilis female preference for the larger L. bibroni males in sympatry, but this requires experimental confirmation. 相似文献
Although waist circumference (WC) is a marker of visceral adipose tissue (VAT), WC cut‐points are based on BMI category. We compared WC‐BMI and WC‐VAT relationships in blacks and whites. Combining data from five studies, BMI and WC were measured in 1,409 premenopausal women (148 white South Africans, 607 African‐Americans, 186 black South Africans, 445 West Africans, 23 black Africans living in United States). In three of five studies, participants had VAT measured by computerized tomography (n = 456). Compared to whites, blacks had higher BMI (29.6 ± 7.6 (mean ± s.d.) vs. 27.6 ± 6.6 kg/m2, P = 0.001), similar WC (92 ± 16 vs. 90 ± 15 cm, P = 0.27) and lower VAT (64 ± 42 vs. 101 ± 59 cm2, P < 0.001). The WC‐BMI relationship did not differ by race (blacks: β (s.e.) WC = 0.42 (.01), whites: β (s.e.) WC = 0.40 (0.01), P = 0.73). The WC‐VAT relationship was different in blacks and whites (blacks: β (s.e.) WC = 1.38 (0.11), whites: β (s.e.) WC = 3.18 (0.21), P < 0.001). Whites had a greater increase in VAT per unit increase in WC. WC‐BMI and WC‐VAT relationships did not differ among black populations. As WC‐BMI relationship did not differ by race, the same BMI‐based WC guidelines may be appropriate for black and white women. However, if WC is defined by VAT, race‐specific WC thresholds are required. 相似文献