分子进化生物学中序列分析方法的新进展

简要介绍了分子进化生物学中序列分析方法的最新进展,特别强调了似然比检验和贝叶斯推论在分子进化和系统发育假说检验中的重要性,并介绍了新方法的一些成功应用,同时还给出了一些重要的信息资源。     

杂交水稻及其“三系”线粒体DNA的AP—PCR指纹图谱

为了研究水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）细胞质雄性不育（ＣＭＳ）与线粒体基因组的关系,应用ＡＰ－ＰＣＲ 分析,用７ 个任意单引物对６ 种水稻品系线粒体ＤＮＡ 进行了扩增。水稻线粒体ＤＮＡ 的ＡＰ－ＰＣＲ 产物可分为三种类型：（１）所有供试品系均能扩增的片段,它们代表了线粒体ＤＮＡ 在进化上的保守性序列。有４ 个引物检测到这类片段。（２）２ 个以上水稻品系共同出现而在全部供试材料间存在差异的扩增片段,这类片段是检测水稻线粒体ＤＮＡ多态性的主要来源。（３）一种细胞质类型所特有的扩增片段,从引物Ｒ２ 和Ｖ５ 的扩增产物中发现了这类片段,它们可能与ＣＭＳ有关联。另外,ＷＡ型不育系珍汕９７Ａ 与其杂种之间在６ 个引物的扩增图谱上均存在不同程度的差异,说明两者的线粒体ＤＮＡ序列结构可能存在某种差别     

HCV E2蛋白诱导的体液免疫及CTL应答研究

应用PCR方法扩增出HCVE2基因编码417a.a～750a.a的DNA片段,克隆到原核表达载体pQE30 LacZ启动子下游,转化JM109菌株.在JM109菌株中诱导表达出N端含6个组氨酸的E2融合蛋白,用Ni-NTA-Superflow亲和层析柱纯化作为抗原免疫实验兔和BALB/c鼠.定期取免血,采用间接ELISA方法检测兔子体内针对E2的抗体水平和维持规律.结果显示,距初次免疫14d兔子体内已有抗体产生,直至免疫第55d抗体水平持续上升,之后抗体水平保持稳定,抗体滴度达到13200.六周后,取鼠脾脏制备淋巴细胞,定向刺激扩增后与经过重组真核表达质粒pCE2转染的P815细胞作用,利用LDH释放试验检测作用效果.在ET=2001的情况下,杀伤率超过30%.这些结果表明工程菌株表达的HCV E2蛋白具有良好的免疫原性,可以诱发免疫实验动物机体产生较高滴度的抗体及特异性CTL应答.由此我们认为E2蛋白是发展HCV预防工程蛋白疫苗的合适候选者.     

Genetic map of Triticum turgidum based on a hexaploid wheat population without genetic recombination for D genome

ABSTRACT: BACKGROUND: A synthetic doubled-haploid hexaploid wheat population, SynDH1, derived from the spontaneous chromosome doubling of triploid F1 hybrid plants obtained from the cross of hybrids Triticum turgidum ssp. durum line Langdon (LDN) and ssp. turgidum line AS313, with Aegilops tauschii ssp. tauschii accession AS60, was previously constructed. SynDH1 is a tetraploidization-hexaploid doubled haploid (DH) population because it contains recombinant A and B chromosomes from two different T. turgidum genotypes, while all the D chromosomes from Ae. tauschii are homogenous across the whole population. This paper reports the construction of a genetic map using this population. RESULTS: Of the 606 markers used to assemble the genetic map, 588 (97%) were assigned to linkage groups. These included 513 Diversity Arrays Technology (DArT) markers, 72 simple sequence repeat (SSR), one insertion site-based polymorphism (ISBP), and two high-molecular-weight glutenin subunit (HMW-GS) markers. These markers were assigned to the 14 chromosomes, covering 2048.79 cM, with a mean distance of 3.48 cM between adjacent markers. This map showed good coverage of the A and B genome chromosomes, apart from 3A, 5A, 6A, and 4B. Compared with previously reported maps, most shared markers showed highly consistent orders. This map was successfully used to identify five quantitative trait loci (QTL), including two for spikelet number on chromosomes 7A and 5B, two for spike length on 7A and 3B, and one for 1000-grain weight on 4B. However, differences in crossability QTL between the two T. turgidum parents may explain the segregation distortion regions on chromosomes 1A, 3B, and 6B. CONCLUSIONS: A genetic map of T. turgidum including 588 markers was constructed using a synthetic doubled haploid (SynDH) hexaploid wheat population. Five QTLs for three agronomic traits were identified from this population. However, more markers are needed to increase the density and resolution of this map in the future study.     

Water Supply Changes N and P Conservation in a Perennial Grass Leymus chinensis

Changes in precipitation can influence soil water and nutrient availability, and thus affect plant nutrient conservation strategies. Better understanding of how nutrient conservation changes with variations in water availability is crucial for predicting the potential influence of global climate change on plant nutrient-use strategy. Here, green-leaf nitrogen (N) and phosphorus (P) concentrations, N- and P-resorption proficiency (the terminal N and P concentration in senescent leaves, NRP and PRP, respectively), and N- and P-resorption efficiency (the proportional N and P withdrawn from senescent leaves prior to abscission, NRE and PRE, respectively) of Leymus chinensis (Trin.) Tzvel., a typical perennial grass species in northern China, were examined along a water supply gradient to explore how plant nutrient conservation responds to water change. Increasing water supply at low levels (< 9000 mL/year) increased NRP, PRP and PRE, but decreased green-leaf N concentration. It did not significantly affect green-leaf P concentration or NRE. By contrast, all N and P conservation indicators were not significantly influenced at high water supply levels (> 9000 mL/year). These results indicated that changes in water availability at low levels could affect leaf-level nutrient characteristics, especially for the species in semiarid ecosystems. Therefore, global changes in precipitation may pose effects on plant nutrient economy, and thus on nutrient cycling in the plant-soil systems.     

贵州黔西县少数民族ABO血型分布及基因频率调查

对贵州黔西县1260例6个少数民族人群红细胞ABO表现型进行了检测;结果如下:贵州黔西县布依族、满族、苗族、白族这四个民族的ABO血型基因频率很相近,彝族和仡佬族与这4个民族的差别较大,布依族,苗族,满族,白族ABO血型分布为O>B>A>AB,彝族为O>A>B>AB,仡佬族为A>O>B>AB;经Hardy Weinberg吻合度检测可以证明贵州黔西县的ABO血型表现型分布状况及基因频率相对稳定,其分布符合hardy Weinberg平衡,获得了该地ABO血型系统群体遗传学数据,为群体遗传学的研究提供了一定的资料。     

不同预处理方法影响昆虫干标本DNA提取效果

为从昆虫干标本中获取高质量DNA,以利于后期PCR扩增目的基因,本研究对DNA提取前昆虫干标本的预处理方法进行探讨,分别测定其纯度和质量,并用不同引物进行了扩增效果对比。结果显示:经过预处理的样品DNA平均含量均高于CK的52.283 ng/滋L,最高为0.9%Na Cl处理3 h,平均含量达122.632 ng/滋L;米象4种预处理方法提取的DNA利用引物Lco1490/Hco2198及UEA7/UEA10均能扩增出650 bp长度条带,仅有0.9%Na Cl溶液浸泡3 h处理的米象可通过引物J173/J1331扩增出1 000 bp的长片段条带。研究表明,经过预处理后,一定程度上减少了基因组DNA的断链,主要对样品DNA的质量浓度影响显著;以9%Na Cl溶液3 h预处理后的昆虫干标本,利用磁珠法所提DNA的质量及PCR扩增效果最佳。     

水通道蛋白1-C末端肽段/GST融合蛋白的原核表达(简报)

水通道蛋白(Aquaporin,AQP)是一类选择性高效转运水分子的细胞膜通道蛋白,广泛存在于原核和真核生物细胞的细胞膜上,主要介导自由水分子的被动跨膜转运,对保持细胞内外液环境的稳态平衡起着重要的作用.     

大气CO2浓度升高对稻季土壤中麦秸降解及氮素分趋的影响

利用中国唯一的稻麦轮作FACE(free-air carbon dioxide enrichment,开放式空气CO2浓度增高)试验平台,研究大气CO2浓度升高对稻季土壤中小麦秸秆降解速率及其氮素分趋的影响.试验设置Ambient(目前空气对照)和FACE(Ambient+200 μmol·mol-1)两个CO2浓度以及低氮处理(LN,150 kg·hm-2)和高氮处理(HN,250 kg·hm-2)两个氮肥水平,在稻季之初按标记麦秸/土壤重量比0.3%添加15N标记小麦秸秆,根据水稻生长时期依次采样测定秸秆降解速率,并通过分析土壤全氮、植株全氮及其15N丰度来观察已降解秸秆的氮素分趋情况.结果发现,大气CO2浓度升高对高氮处理土壤中小麦秸秆降解速率没有显著影响,但显著促进了低氮处理土壤中小麦秸秆的降解(p < 0.05),使其提高到与高氮处理土壤相当水平;大气CO2浓度升高显著增加了已降解秸秆中氮素的流失,在高氮处理土壤中尤为严重,而对植物吸收已降解秸秆中的氮素没有显著影响.结果表明,大气CO2浓度升高在土壤氮素相对不足时会加速土壤中小麦秸秆的降解,而在土壤氮素相对充足时又会加大降解秸秆中氮素的流失.     

Language Differences in the Brain Network for Reading in Naturalistic Story Reading and Lexical Decision

Differences in how writing systems represent language raise important questions about whether there could be a universal functional architecture for reading across languages. In order to study potential language differences in the neural networks that support reading skill, we collected fMRI data from readers of alphabetic (English) and morpho-syllabic (Chinese) writing systems during two reading tasks. In one, participants read short stories under conditions that approximate natural reading, and in the other, participants decided whether individual stimuli were real words or not. Prior work comparing these two writing systems has overwhelmingly used meta-linguistic tasks, generally supporting the conclusion that the reading system is organized differently for skilled readers of Chinese and English. We observed that language differences in the reading network were greatly dependent on task. In lexical decision, a pattern consistent with prior research was observed in which the Middle Frontal Gyrus (MFG) and right Fusiform Gyrus (rFFG) were more active for Chinese than for English, whereas the posterior temporal sulcus was more active for English than for Chinese. We found a very different pattern of language effects in a naturalistic reading paradigm, during which significant differences were only observed in visual regions not typically considered specific to the reading network, and the middle temporal gyrus, which is thought to be important for direct mapping of orthography to semantics. Indeed, in areas that are often discussed as supporting distinct cognitive or linguistic functions between the two languages, we observed interaction. Specifically, language differences were most pronounced in MFG and rFFG during the lexical decision task, whereas no language differences were observed in these areas during silent reading of text for comprehension.