粗枝云杉( Picea asperata Mast.)天然群体的遗传变异

西南地区在我国的经济发展和生态环境建设中占重要地位，但也是我国生态环境最脆弱的地区之一，生态系统退化，生态功能减弱，严重制约着西南林业的可持续经营与发展。本项目采用DNA 分子标记SSR 研究不同生境条件下粗枝云杉群体的遗传变异及其时空分布格局，考察遗传变异与复杂的山地生态环境间的潜在联系，系统地揭示粗枝云杉天然群体与环境系统相互作用的生态适应与分子进化机制。粗枝云杉适应性强，生长迅速，在植树造林和工业用材方面占有重要地位，研究成果可为中国西南部亚高山天然林的可持续经营及退化生态系统的恢复与重建提供理论依据和科学指导。主要研究结果如下： 1. SSR 位点变异丰富，等位基因频率的分布格局多样。7 个SSR 标记全是多态位点，每位点的等位基因数变化范围为13~24，平均为19.9 个。SSR 位点的等位基因片段长度范围变化较大。73.1%的等位基因变异遵循逐步突变模型(SSM)而发生1 个重复基元的变化，22.3%和4.6%的变异分别按两阶段突变模型(TMP)发生1 个重复基元以上的变化和在SSR 位点侧翼区发生1 个碱基变化的插入-删除事件。 2. 粗枝云杉拥有中等偏高水平的遗传多样性和相对大的群体间遗传分化。通过分析代表10 个群体的250 个个体在7 个SSR位点的变化，调查了源自中国西南山区的粗枝云杉的微卫星变异。相当高的遗传多样性和强烈的群体分化发生在粗枝云杉中， 其群体平均Nei's 期望杂合度为0.707 ， 群体间遗传距离为0.121~0.224(FST)和0.100~0.537(RST)。然而，群体间遗传距离与地理距离之间无相关性，从而排除了简单的距离分离模式并暗示迁移不是影响粗枝云杉遗传变异格局的主要因素。事实上，使用私有等位基因估算的基因流数量非常低，仅等于0.753。等位基因置换检验(Allele permutation tests)揭示逐步突变及遗传漂变都对群体间分化有贡献。另外，在多数位点检测到显著的群体间遗传差异，这个结果说明自然选择，假设通过环境压力，是引起粗枝云杉微地理分化的主要因素之一。根据SSR基因型，250 个粗枝云杉个体的70%被正确地归类入其各自的来源群体，结果表明微卫星(SSR)对区分来自中国不同生态地理位点的粗枝云杉基因型是有效的。 3. 在SSR、RAPD 和AFLP 位点，显著的群体间遗传结构被发现的，但三种标记间遗传分化程度和群体遗传关系有差异。利用来自10 个群体的247 个个体，我们报告了关于样本粗枝云杉群体间遗传关系的总体看法。根据各自对评价遗传关系的信息能力和适用性，SSR、RAPD 和AFLP 标记被选用，三种技术非常有效地区别这些基因型。使用的SSR、RAPD 和AFLP 标记分别估计平均Dice 相似性系数。Mantel 检验产生显著但相对低的共表型适合度(RAPD = 0.63£AFLP = 0.60和SSR = 0.75)。比较三种标记系统，RAPD 和AFLP 共表型指数相对高地相关(r =0.59)，而RAPD 和SSR 及SSR 和AFLP 之间的相关系数分别是0.53 和0.35。所有系统树，包括不同标记资料结合获得的系统树，反映了多数群体依据它们的地理条件而成某种特定关系。结果暗示单个或结合标记系统能用来深入洞察粗枝云杉遗传研究，并且不同标记系统合并资料能提供更可靠的信息。 Southwestern region plays an important role in economic developmentand ecological construction in China. Yet, it is also one of the weak regionsof ecological environment in China with degraded ecosystem and imperfectfunction, which restricts the sustaining management and development ofsouthwestern forestry. The genetic variation and spatial distribution patternof P. asperata populations originating from different habitats wereinvestigated using SSR molecular markers in this study. The correlationsbetween genetic variation and ecological and environmental conditionswere detected, and the interaction between P. asperata populations andenvironmental system and the mechanism of ecological adaption -molecular evolution were revealed. Given the significant ecological andeconomic roles of the fast-growing and wide-adaptive species in reforestation and production of pulp wood and timber, the study couldprovide a strong theoretical evidence and scientific direction for thesustaining management of subalpine natural forest, and the afforestationand rehabilitation of degraded ecosystem. The results are as follows: 1. The genetic variation at SSR loci was abundant and the distributionof allelic frequencies was uneven. All seven loci were polymorphic, and thenumber of alleles per locus varied from 13 to 24 with a mean valueequaling 19.9. The allele sizes at SSR loci were found to vary widely.73.1% of allelic variation followed stepwise mutation model (SSM) whichresults increase or decrease by one repeat type, and 22.3% and 4.6% wereresulted from two-phase mutation model (TMP) with allele size varying bymore than one repeat type and from insertion-deletion events in theflanking regions at SSR loci with a single basepair changing, respectively. 2. P. asperata possessed a moderate to high level of genetic diversityand considerable genetic differentiation. Microsatellite variation of P.asperata. originating from the mountains of southwestern China wasinvestigated by analyzing variation at seven SSR loci in 250 individualsrepresenting ten populations. A fair degree of genetic diversity and strongpopulation subdivision occurred with the mean gene diversity (H) of 0.707,and genetic distances among populations varying between 0.121 and 0.224(FST) and between 0.100 and 0.537 (RST). However, inter-populationgenetic distances showed no correlation with geographic distances between the population sites. This ruled out a simple isolation by distance modeland suggested that migration does not have a great impact. In fact, theamount of gene flow, detected using private alleles, was very low, equalingonly 0.753. Allele permutation tests revealed that stepwise-like mutations,coupled with genetic drift, could contribute to population differentiation.Moreover, significant genetic differences between populations weredetected at most loci. The results indicate that natural selection, presumablythrough environmental stress, may be one of the main factors causingmicro-geographical differentiation in the genetic structure of P. asperata.Based on SSR genotypes, 70% of the 250 individuals were correctlyclassified into their sites of origin. This suggests that microsatellites (SSRs) are effective in distinguishing genotypes of P. asperata originating fromdiverse eco-geographical sites in China. 3. Using a set of 247 individuals from ten P. asperata populations wereport an overview on the genetic relationship among the sampled P.asperata populations. RAPD, AFLP and SSR were used in terms of theirinformativeness and applicability for evaluate relationship and all threetechniques discriminated the genotypes very effectively. Mean Dicesimilarities coefficient were estimated using RAPD, AFLP and SSR,respectively. The Mantel test resulted in a significant but relatively low fit(RAPD = 0.63, AFLP = 0.60 and SSR = 0.75) of cophenetic values.Comparing the three marker systems to each other, RAPD and AFLP cophenetic indices were highly correlated (r = 0.59), while correlationcoefficient between RAPD and SSR was r = 0.53 and between SSR andAFLP was r = 0.35. For all markers a relatively high similarity indendrogram topologies was obtained although some differences wereobserved. All the dendrograms, including that obtained by the combineduse of all the marker data, reflect some relationships for most of thepopulations according to their geographic conditions. The results indicatethat single or combined marker system could be used to insight into geneticstudy in P. asperata and the combined data of different marker systems canprovide more reliable information.

PE文件隐型加壳技术的研究与实现

通过对PE(portable executable)文件格式的了解,编写PE分析工具对文件内部结构进行分析。详细介绍了对PE可执行文件加壳的全过程,在此过程中巧妙地使用MD5、CRC32等成熟的hash算法及防API断点跟踪等多种反破解技术,并采用自动隐藏加密方案,大大地提高了软件的保护力度。

高LET重离子照射人肿瘤细胞的DNA双链断裂及修复研究

依托兰州近代物理研究所的HIRFL装置对人肝癌细胞SMMC-7721进行高LET的重离子辐照,同时利用脉冲场凝胶电泳研究高LET辐射引起的细胞DSB及其修复的特点。对于不同离子的辐射进行了比较分析,探索重离子的辐射生物学效应机理。结果发现,随着剂量的增加,初始DSB在给定剂量范围内呈线性增长,LET较高的氩离子的剂量效应曲线具有较大的斜率。对于DSB分布情况的研究发现,LET不同,大小片段的分布随着剂量的变化呈现不同的变化情况。修复研究显示两种LET辐射的快修复在30min左右都已经结束,此后都开始了慢修复,对于高LET的氩离子辐射来说,快修复和慢修复完成的修复量均高于较低LET的碳离子辐射。4h时,两种辐射残余的DSB基本一致。结果证明了高LET辐射下初始DSB与剂量的线性依赖关系,且发现不同种类的离子辐射引起的DSB的剂量效应曲线的斜率不同。结果也显示了DSB的非随机分布现象和DSB修复的双阶段模式,发现LET、离子种类和剂量对于DSB的片段非随机分布同时存在影响,同时修复结果提示慢修复机制和错修复在高LET辐射引起损伤的修复研究中可能具有重要意义。

高能重离子辐照制备碳氮化合物材料研究

室温下,先用100—120keV的N离子注入类金刚石薄膜和石墨中,注入剂量5×1017至5×1018 cm-2,再用高能Xe、U、C60离子分别辐照注氮后的样品,然后用显微FTIR和Raman、XRD/XPS等手段进行分析表征,研究了实验样品中由辐照引起的新化学键和新相的产生。实验结果显示,高能重离子辐照可在所有样品中产生大量的CN键,高N浓度和大密度能量沉积导致sp3/sp2键比率的增加以及形成α-和β-C3N4必需的N-sp3C键的量的增加。C60离子辐照在注氮石墨样品中引起了ta-C、N=sp2C和N-sp3C键的形成;而高能离子辐照在注氮类金刚石薄膜样品中产生了α-和β-C3N4晶态夹杂物,其尺寸在1.4—3.6nm之间。

γ射线对A172胶质瘤细胞的生物学效应

以不同剂量照射后的细胞存活率、微核率和微核细胞率作为生物学终点,研究了γ射线对A172细胞的生物学效应.结果表明:细胞存活率与剂量之间满足回归方程lgY=-0.06427X+1.83354,其回归系数r=-0.9886,P<0.01.剂量为1Gy时微核率和微核细胞率达到最大值,此时的微核率为(66.75±3.564)%,微核细胞率为(53.9±0.7849)%,微核率和微核细胞率均随着剂量的增大先增大后减小,并分别维持在42%和37%左右.

单核子在具有八极形变的平均势场中的混沌运动(Ⅲ)初始相干态空时演化中几率密度及扰动强度的研究

运用在二维不对称谐振子势加八极形变势中传播的二维不对称谐振子相 干态，研究了不同强度耦合作用下密度矩阵的复杂性及对干扰动强度微小变动 的敏感性。发现相应于对应经典系统作规则、部分混沌、及整体混沌运动的相空 间区域，量子系统有对应的特征性的表现，它们与势能面上负曲率的存在及负 曲率的大小有关系．

Study on DNA Damage Induced by Neon Beam Irradiation in Saccharomyces Cerevisiae

Yeast strain Saccharornyces cerevisiae was irradiated with different doses of 85 MeV/u Ne-20(10+) to investigate DNA damage induced by heavy ion beam in eukaryotic microorganism. The survival rate, DNA double strand breaks (DSBs) and DNA polymorphic were tested after irradiation. The results showed that there were substantial differences in DNA between the control and irradiated samples. At the dose of 40 Cy, the yeast cell survival rate approached 50%, DNA double-strand breaks were barely detectable, and significant DNA polymorphism was observed. The alcohol dehydrogenase II gene was amplified and sequenced. It was observed that base changes in the mutant were mainly transversions of T-->G and T-->C. It can be concluded that heavy ion beam irradiation can lead to change in single gene and may be an effective way to induce mutation.