中国鄂温克族和达斡尔族人群中血小板膜糖蛋白Ib(GPIb)的多态性

研究旨在进一步探索同一人种的不同民族之间GPIb的多态性是否存在差异。血小板从84名健康鄂温克族,85名达斡尔族青年志愿采血分离而获得,血小板样品经SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）后,再转移至硝酸纤维素膜上（Western blotting),用生物素化的素胚凝集素（WGA/Bio)偶联辣根过氧化酶复合物试剂盒（ABCkit)分步孵育,再以4-氯-1-萘酚显影,膜上呈现的深灰色带即是GPIbα链,其分子量分别为141、136、132、128kD,此即A、B、C、D四型,此为基因型,其中每两型构成一个表型,本研究证明,在我国鄂温克族人群中,等位基因A、B、C、D的频率分别为0.113,0.375,0.381,0.131;在达斡尔族中,A型为0.124,B型为0.406,C型为0.359。D型为0.112。经统计学分析。在这两个民族的人群之间,其GPIb各基因频率无显性差异。两个民族人群的GPIb表型经卡方检验也无显性差异。由此结果说明,鄂温克,达斡尔两个民族的人群之间,GPIb的多态性特征是相似的。     

Robust Estimates of Divergence Times and Selection with a Poisson Random Field Model: A Case Study of Comparative Phylogeographic Data

Mutation frequencies can be modeled as a Poisson random field (PRF) to estimate speciation times and the degree of selection on newly arisen mutations. This approach provides a quantitative theory for comparing intraspecific polymorphism with interspecific divergence in the presence of selection and can be used to estimate population genetic parameters. Although the original PRF model has been extended to more general biological settings to make statistical inference about selection and divergence among model organisms, it has not been incorporated into phylogeographic studies that focus on estimating population genetic parameters for nonmodel organisms. Here, we modified a recently developed time-dependent PRF model to independently estimate genetic parameters from a nuclear and mitochondrial DNA data set of 22 sister pairs of birds that have diverged across a biogeographic barrier. We found that species that inhabit humid habitats had more recent divergence times and larger effective population sizes than those that inhabit drier habitats, and divergence time estimated from the PRF model were similar to estimates from a coalescent species-tree approach. Selection coefficients were higher in sister pairs that inhabited drier habitats than in those in humid habitats, but overall the mitochondrial DNA was under weak selection. Our study indicates that PRF models are useful for estimating various population genetic parameters and serve as a framework for incorporating estimates of selection into comparative phylogeographic studies.     

珍稀濒危植物珙桐胚的萌发与快速繁殖

首次采用均匀设计和回归分析方法对影响珙桐(Davidia involucrata)胚萌发的3种因素(基本培养基、赤霉素和活性炭)进行优化, 并利用萌发胚建立了珙桐的高效再生体系。 利用回归方程得出胚萌发的最优条件为: 在含有0.5 mg.L^{- 1} IBA和1 mg.L^-1 6-BA的1/2MS培养基中添加2.85 mg.L^-1 GA3和0.25 g.L^-1活性炭。在验证实验中珙桐胚萌发率为87.72%, 与预测值无显著差异。 最优的芽诱导与增殖条件为: 添加0.1 mg.L^-1 NAA和2.0 mg.L^-1 6-BA 的WPM培养基, 增殖系数为4.34。在生根培养基中增殖的芽生根率高达90%, 移栽成活率为71%。     

Evaluating the role of contracting and expanding rainforest in initiating cycles of speciation across the Isthmus of Panama

Climatic and geological changes across time are presumed to have shaped the rich biodiversity of tropical regions. However, the impact climatic drying and subsequent tropical rainforest contraction had on speciation has been controversial because of inconsistent palaeoecological and genetic data. Despite the strong interest in examining the role of climatic change on speciation in the Neotropics there has been few comparative studies, particularly, those that include non-rainforest taxa. We used bird species that inhabit humid or dry habitats that dispersed across the Panamanian Isthmus to characterize temporal and spatial patterns of speciation across this barrier. Here, we show that these two assemblages of birds exhibit temporally different speciation time patterns that supports multiple cycles of speciation. Evidence for these cycles is further corroborated by the finding that both assemblages consist of 'young' and 'old' species, despite dry habitat species pairs being geographically more distant than pairs of humid habitat species. The matrix of humid and dry habitats in the tropics not only allows for the maintenance of high species richness, but additionally this study suggests that these environments may have promoted speciation. We conclude that differentially expanding and contracting distributions of dry and humid habitats was probably an important contributor to speciation in the tropics.     

Statistical inference of selection and divergence from a time-dependent Poisson random field model

We apply a recently developed time-dependent Poisson random field model to aligned DNA sequences from two related biological species to estimate selection coefficients and divergence time. We use Markov chain Monte Carlo methods to estimate species divergence time and selection coefficients for each locus. The model assumes that the selective effects of non-synonymous mutations are normally distributed across genetic loci but constant within loci, and synonymous mutations are selectively neutral. In contrast with previous models, we do not assume that the individual species are at population equilibrium after divergence. Using a data set of 91 genes in two Drosophila species, D. melanogaster and D. simulans, we estimate the species divergence time t(div) = 2.16 N(e) (or 1.68 million years, assuming the haploid effective population size N(e) = 6.45 x 10(5) years) and a mean selection coefficient per generation μ(γ) = 1.98/N(e). Although the average selection coefficient is positive, the magnitude of the selection is quite small. Results from numerical simulations are also presented as an accuracy check for the time-dependent model.     

血浆糖蛋白β2-GPI分子结构域与ox-LDL结合机制

通过酵母重组P.rβ_2-GPI-DV(β2糖蛋白I第5结构域)蛋白的荧光、圆二色谱及分子对接模拟,分析血浆糖蛋白β2-GPI-DV分子结构域与ox-LDL的结合机制。SDS-PAGE、Western blotting验证重组蛋白P.rβ_2-GPI-DV表达正确性及纯度;荧光、圆二色光谱、分子对接模拟解析重组蛋白与oxLDL、oxLDL配体oxLig-1可能存在的结合机制和位点。SDS-PAGE、Western blotting显示,在12 kDa大小处有目的蛋白存在且与特异性抗体结合;荧光、圆二色谱的发色基团、二级结构的变化趋势及分子对接模拟结果揭示,P.rβ_2-GPI-DV的Cys281-Lys-Asn-Lys-Glu-Lys-Lys287和Leu313-Ala-Phe-Trp316区域组成的活性口袋与oxLig-1的ω-COOH羧基是实现二者结合的关键。以上结果表明,β2-GPI通过其第5结构域(DV)的赖氨酸阳性区域与ox-LDL的ω-COOH基团识别结合,为血清中β2-GPI特异性结合ox-LDL的机制研究提供理论依据。     

珍稀濒危植物珙桐胚的萌发与快速繁殖

首次采用均匀设计和回归分析方法对影响珙桐（Davidia involucrata）胚萌发的3种因素（基本培养基、赤霉素和活性炭）进行优化,并利用萌发胚建立了珙桐的高效再生体系。利用回归方程得出胚萌发的最优条件为：在含有0．5mg·L^-1 IBA和1mg·L^-16·BA的1/2MS培养基中添加2．85mg·L^-1GA3和0．25g·L^-1活性炭。在验证实验中珙桐胚萌发率为87．72％,与预测值无显著差异。最优的芽诱导与增殖条件为：添加0．1mg·L^-1NAA和2．0mg·L^-16-BA的WPM培养基,增殖系数为4．34。在生根培养基中增殖的芽生根率高达90％,移栽成活率为71％。     

Proteomic identification of intracellular vesicle trafficking and protein glycosylation requirements for lumen inflation in Ciona notochord

Lumen formation and inflation are crucial steps for tubular organ morphogenesis, yet the underling mechanism remains largely unrevealed. Here, we applied 4D proteomics to screen the lumenogenesis-related proteins and revealed the biological pathways potentially that are involved in lumen inflation during notochord lumen formation in the ascidian Ciona savignyi. In total, 910 differentiated expressed proteins (DEPs) were identified before and after notochord lumen formation utilizing Mfuzz analysis. Those DEPs were grouped into four upregulated clusters based on their quantitative expression patterns; the functions of these proteins were enriched in protein metabolic and biosynthetic process, the establishment of localization, and vesicle-mediated transport. We analyzed the vesicle trafficking cluster and focused on several vesicle transport hub proteins. In vivo function-deficient experiments showed that mutation of vesicle transport proteins resulted in an abnormal lumen in notochord development, demonstrating the crucial role of intracellular trafficking for lumen formation. Moreover, abundant extracellular matrix proteins were identified, the majority of which were predicted to be glycosylated proteins. Inhibition of glycosylation markedly reduced the lumen expansion rate in notochord cells, suggesting that protein glycosylation is essential for lumenogenesis. Overall, our study provides an invaluable resource and reveals the crucial mechanisms in lumen formation and expansion.