落叶松属的分子进化与生物地理学

　　松科植物的核基因组十分庞大，基因常形成复杂的基因家族，核rDNA ITS 区在基因组内和基因组间存在广泛的长度和序列变异，但染色体数目和核型却高度保守，几乎均为二倍体(2n＝24)，与被子植物频繁的多倍化和高度均一的ITS区形成鲜明对比;叶绿体、线粒体和核基因组分别为父系、母系及双亲遗传，这种独特的遗传体系组合为系统发育重建研究提供了便利条件。因此，松科植物不仅是阐明基因树/物种树这一理论问题的理想试材，而且是基因和基因组进化及核rDNA致同进化机制研究的好材料。此外，松科植物的进化历史悠久，很多类群经历了多次重大的地质历史事件，并呈各种间断分布格局，其生物地理学问题受到广泛关注。本文对落叶松属所有物种(L. lyallii除外)和大部分变种的叶绿体基因组trnT-trnF区、低拷贝核4CL基因家族 (4-香豆酸辅酶A连接酶基因)及多拷贝核rDNA ITS区进行了序列分析，重建了该属的系统发育并揭示了其地理分布格局的形成过程，同时基于克隆和基因谱系分析，探讨了核4CL和rDNA ITS这两个基因家族的进化式样及规律。 　　1. 叶绿体trnT-trnF区和核rDNA ITS区的研究结果表明：落叶松属的种间遗传分化程度很低，北美的种类构成一个单系分支，并为欧亚种类的姐妹群。短苞鳞的欧亚落叶松组和长苞鳞的欧亚红杉组之间的分化较早，接近欧亚和北美种类间的分化时间。换句话说，苞鳞长短的分化在落叶松属中至少发生过两次，其中一次在落叶松属分化的初期，另一次在北美的种类中。结合化石、地史及气候资料，我们推测：落叶松属的共同祖先通过白令陆桥扩散，并形成欧亚和北美两支,然后在不同的板块上独立进化。落叶松组的泛北极分布是冰期后的回迁形成的，而红杉组的物种在第三纪全球气温降低时向南迁移，进而形成东亚－北美间断分布，特别是欧亚红杉组的祖先曾伴随青藏高原的隆升而发生辐射分化。 　　2. 在落叶松属4CL基因家族的研究中共获得44个差异的克隆，除华北落叶松外，其它种类均含2-4个成员。系统发育分析表明： 4CL基因频繁发生重复/丢失，并导致谱系拣选。该基因在落叶松属的共同祖先中发生一次重复，形成4clA和4clB，4clA再次发生基因重复形成4clA1和4clA2。重复产生的这两对并系基因拷贝在进化速率上呈显著差异，其中一个拷贝的进化速率明显加快，可能与进化制约的减弱或功能分化有关。结合其它核基因的研究结果，我们推测频繁的基因重复/丢失可能是形成和维持松科植物庞大核基因组的重要机制之一。 　　3. 对落叶松属101个nrDNA ITS克隆进行了序列及分子进化分析，发现极少数克隆存在较大的长度及（或）序列变异，并可能为假基因或重组体，其它克隆间的序列分化水平较低。因而，落叶松属核rDNA的致同进化速率比松科中两个古老的属（松属和云杉属）快。该致同进化速率的加快可能与落叶松属年轻的进化历史及染色体上较少的rDNA位点数目有关。由于一些特异克隆含嵌合序列及极高的序列变异，推测它们可能来源于物种进化过程中染色体重排形成的小位点(minor loci)或为孤独基因(orphons)。此外，我们发现nrDNA ITS克隆的分布式样与落叶松属的分化及地理分布格局的形成有密切关系：在欧亚红杉组中，克隆常按分类群（物种或变种）形成单系分支，表明这些类群的分化曾伴随着强烈的nrDNA ITS奠基者效应;相反，在欧亚落叶松组中，所有物种的克隆均混杂在一起，说明这些物种的分化时间较晚或在冰期后回迁的过程中曾发生频繁的种间基因交流。

火烧、施肥、刈割及其交互作用对草原土壤净氮矿化的影响

我国北方温带草原地处干旱半干旱区，是欧亚大陆草原生物区系的重要组成部分，也是我国重要的畜牧业生产基地。土壤氮素作为陆地植物生长和生态系统初级生产力的主要限制因子之一，了解其矿化和可利用性对各种人为干扰因素的响应，有助于我们充分认识草地退化机理，维持草场生产力并进一步促进畜牧业的可持续发展，为草地恢复、重建和维护提供理论依据。本实验在中国科学院内蒙古草原生态系统定位研究站的多年围封退化样地，分别实施了火烧、施肥、刈割及其交互作用的人为干扰处理，利用原状土野外培养的方法，在两年时间中，研究不同的人为干扰因素（火烧、施肥、刈割及其交互处理）对土壤净氮矿化作用的影响。 火烧显著影响草原的氮循环过程，季节和年际的气候变化参与调节氮循环对火烧处理的响应。多年围封后的初次人工火烧处理显著降低了第一个生长季（2006 年生长季）和其后冬季的土壤净氮矿化速率，但是，火烧处理仅在冬季对土壤无机氮含量产生显著降低作用；2007 年生长季，火烧处理对土壤净氮矿化速率和土壤无机氮含量没有显著作用，但是在个别月份，不同的火烧频率对土壤净氮矿化速率和无机氮含量的影响表现出差异。2006 年生长季，未火烧样地和火烧样地净氮矿化积累量分别为3.07±0.26 g N m-2 和2.18±0.21 g N m-2；冬季，未火烧样地和火烧样地净氮矿化积累量分别为1.18±0.25 g N m-2 和0.51±0.08 g N m-2；2007 年生长季，未火烧样地（BC）、每年火烧样地（B1）和两年一烧样地（B2）净氮矿化积累量分别为1.32±0.21 g N m-2、0.54±0.30 g N m-2 和 0.77±0.24 g N m-2。较为湿润的2006 年生长季的净氮矿化积累量显著高于较为干旱的2007 年生长季，冬季也存在相当丰富的净氮矿化积累量。我们推论，长时间围封后的单次火烧处理对土壤净氮矿化作用的影响可能是短效的，但是年际间气候变化对土壤净氮矿化作用影响显著。 施肥显著提高了土壤的无机氮含量，并且与施肥梯度呈显著正相关关系。施肥对土壤无机氮含量的影响具有累加效应，第一次施肥和第二次施肥后的首次取样，+N5.25 、+N17.5 、+N28.0 三个施肥梯度的样地土壤无机氮含量比未施肥样地分别高出56%、219%、1054%和514%、891%、1811%。施肥处理在2006 年和2007 年对土壤净氮矿化作用都没有显著影响，仅在2007 年的个别月份有一定效果。以上结果说明，无机氮肥的添加可以明显提高土壤中无机氮的含量，满足植物生长的需求，但对于土壤氮转化过程的影响可能还要受其它环境和生物因子的制约。 刈割对土壤无机氮含量和净硝化速率没有显著作用，对土壤净氮矿化速率仅在个别月份表现显著效果。2007 年生长季，未刈割样地和刈割样地净氮矿化积累量分别为1.32±0.21 g N m-2 和1.08±0.35 g N m-2，不存在显著差异。我们推论，长期围封后的单次刈割处理在短期内对生长季的土壤氮动态仅有微弱影响，并且这种效果还可能受水分因素的制约。 火烧、施肥、刈割的交互处理在2007 年生长季对土壤无机氮含量产生显著作用，但是，对土壤净氮矿化速率和净硝化速率没有显著效果，然而，在不同的取样时间，火烧、施肥、刈割的交互处理对土壤无机氮含量、净氮矿化速率和净硝化速率的影响存在显著差异，说明火烧、施肥、刈割的交互处理对土壤无机氮和净氮矿化作用的影响可能受各种环境因子的制约，尤其是在水分相对缺乏的半干旱内蒙古草原，非生物因子和生物因子可能共同影响着土壤中的氮平衡。 本研究初步分析了长期围封后火烧、施肥、刈割及其交互处理对土壤净氮矿化的影响，初步探究了人为干扰和环境气候变化对土壤可利用性氮的调节作用，为科学地进行禁牧、割草、人工养份添加等草原管理提供了数据支持。

Effects of morphine and its withdrawal on Y-maze spatial recognition memory in mice

Effects of morphine on acquisition and retrieval of memory have been proven in the avoidance paradigms. In present study, we used a two-trial recognition Y-maze to test the effects of acute morphine and morphine withdrawal on spatial recognition memory. T

VISUALIZATION OF THE CHROMOSOME SCAFFOLD IN A PRIMITIVE EUKARYOTE DINOFLAGELLATE CRYPTHECOAINIUM-COHNII

The chromosome scaffolds in higher eukaryotic nuclei have been described elsewhere. But it is unknown when they evolved. The dinoflagellates are the primitive organisms that may be the intermediate between prokaryotes and eukaryotes. Combining chromosome scaffold preparation methods with embedment-free section microscopy, we demonstrate that the dinoflagellate Crypthecodinium cohnii chromosome retains a protein scaffold after the depletion of DNA and soluble proteins. This scaffold preserves the morphology characteristic of the chromosome. Two-dimensional electrophoreses show that the chromosome scaffolds are mainly composed of acidic proteins. Our results suggest that a framework similar to the chromosome scaffold in the mammalian cell appeared in the primitive eukaryote. We propose that the chromosome scaffold possibly originated from the early stages of eukaryote evolution.

Isolation and cloning of a metalloproteinase from king cobra snake venom

A 50 kDa fibrinogenolytic protease, ohagin, from the venom of Ophiophagus hannah was isolated by a combination of gel filtration, ion-exchange and heparin affinity chromatography. Ohagin specifically degraded the alpha-chain of human fibrinogen and the pr

Molecular characterization of L-amino acid oxidase from king cobra venom

An L-amino acid oxidase from Ophiophagus hannah snake venom (Oh-LAAO) was purified by successive gel filtration, ion-exchange and heparin chromatography. Oh-LAAO did not induce platelet aggregation; however, it had potent inhibitory activity on platelet a

Diffusive coevolution and mutualism maintenance mechanisms in a fig-fig wasp system

In reciprocal mutualism systems, the exploitation events by exploiters might disrupt the reciprocal mutualism, wherein one exploiter species might even exclude other coexisting exploiter species over an evolutionary time frame. What remains unclear is how such a community is maintained. Niche partitioning, or spatial heterogeneity among the mutualists and exploiters, is generally believed to enable stability within a mutualistic system. However, our examination of a reciprocal mutualism between a fig species (Ficus racemosa) and its pollinator wasp (Ceratosolen fusciceps) shows that spatial niche partitioning does not sufficiently prevent exploiters from overexploiting the common resource (i.e., the female flowers), because of the considerable niche overlap between the mutualists and exploiters. In response to an exploiter, our experiment shows that the fig can (1) abort syconia-containing flowers that have been galled by the exploiter, Apocryptophagus testacea, which oviposits before the pollinators do; and (2) retain syconia-containing flowers galled by Apocryptophagus mayri, which oviposit later than pollinators. However, as a result of (2), there is decreased development of adult non-pollinators or pollinator species in syconia that have not been sufficiently pollinated, but not aborted. Such discriminative abortion of figs or reduction in offspring development of exploiters while rewarding cooperative individuals with higher offspring development by the fig will increase the fitness of cooperative pollinating wasps, but decrease the fitness of exploiters. The fig fig wasp interactions are diffusively coevolved, a case in which fig wasps diversify their genotype, phenotype, or behavior as a result of competition between wasps, while figs diverge their strategies to facilitate the evolution of cooperative fig waps or lessen the detrimental behavior by associated fig wasps. In habitats or syconia that suffer overexploitation, discriminative abortion of figs or reduction in the offspring development of exploiters in syconia that are not or not sufficiently pollinated will decrease exploiter fitness and perhaps even drive the population of exploiters to local extinction, enabling the evolution and maintenance of cooperative pollinators through the movement between habitats or syconia (i.e., the metapopulations).

Xenopus RCOR2 (REST corepressor 2) interacts with ZMYND8, which is involved in neural differentiation

Regulation of neuronal gene expression is critical to nervous system development. REST (RE1-silencing transcription factor) regulates neuronal gene expression through interacting with a group of corepressor proteins including REST corepressors (RCOR). Here we show that Xenopus RCOR2 is predominantly expressed in the developing nervous system. Through a yeast two-hybrid screen, we isolated Xenopus ZMYND8 (Zinc finger and MYND domain containing 8) as an XRCOR2 interacting factor. XRCOR2 and XZMYND8 bind each other in co-immunoprecipitation assays and both of them can function as transcriptional repressors. XZMYND8 is co-expressed with XRCOR2 in the nervous system and overexpression of XZMYND8 inhibits neural differentiation in Xenopus embryos. These data reveal a RCOR2/ZMYND8 complex which might be involved in the regulation of neural differentiation. (C) 2010 Elsevier Inc. All rights reserved.

RNF220, an E3 ubiquitin ligase that targets Sin3B for ubiquitination

Modification of proteins by ubiquitination plays important roles in various cellular processes. During this process, the target specificity is determined by ubiquitin ligases. Here we identify RNF220 (RING finger protein 220) as a novel ubiquitin ligase for Sin3B. As a conserved RING protein, RNF220 can bind E2 and mediate auto-ubiquitination of itself. Through a yeast two-hybrid screen, we isolated Sin3B as one of its targets, which is a scaffold protein of the Sin3/HDAC (histone deacetylase) corepressor complex. RNF220 specifically interacts with Sin3B both in vitro and in vivo. Sin3B can be regulated by the ubiquitin-proteasome system. Co-expression of RNF220 promotes the ubiquitination and proteasomal degradation of Sin3B. Taken together, these results reveal a new mechanism for regulating the Sin3/HDAC complex. (C) 2010 Elsevier Inc. All rights reserved.

A phylogeny of Chinese species in the genus Phrynocephalus (Agamidae) inferred from mitochondrial DNA sequences

We investigated the phylogenetic relationships among most Chinese species of lizards in the genus Phrynocephalus (118 individuals, collected from 56 populations of 14 well-defined species and several unidentified specimens) using four mitochondrial gene fragments (12S rRNA, 16S rRNA, cytochrome b, and ND4-tRNA(LEU)). The partition-homogeneity tests indicated that the combined dataset was homogeneous, and maximum-parsimony (MP), neighbor-joining (NJ), maximum-likelihood (ML) and Bayesian (BI) analyses were performed on this combined dataset (49 haplotypes including outgroups for 2058 bp in total). The maximum-parsimony analysis resulted in 24 equally parsimonious trees, and their strict consensus tree shows that there are two major clades representing the Chinese Phrynocephalus species: the viviparous group (Clade A) and the oviparous group (Clade B). The trees derived from Bayesian, ML. and NJ analyses were topologically identical to the MP analysis except for the position of P. mystaceus. All analyses left the nodes for the oviparous group, the most basal clade within the oviparous group, and P. mystaceus unresolved. The phylogenies further suggest that the monophyly of the viviparous species may have resulted from vicariance, while recent dispersal may have been important in generating the pattern of variation among the oviparous species. (C) 2003 Elsevier Science (USA). All rights reserved.

Origin of rabbit (Oryctolagus cuniculus) in China: evidence from mitochondrial DNA control region sequence analysis

A fragment of mitochondrial DNA (mtDNA) control region (similar to700 bp) was sequenced in 104 individuals from 20 breeds (three Chinese domestic breeds, five recently derived breeds and 12 introduced breeds) of domestic rabbits, Oryctolagus cuniculus . Nineteen sites were polymorphic, with 18 transitions and one insertion/deletion, and eight haplotypes (A1, A2, A3, A4, A5, A6, A7 and A8) were identified. Haplotype A1 was the most common and occurred in 89 individuals. In the 25 Chinese rabbits, only haplotype A1 was observed, while four haplotypes (A1, A3, A5 and A6) were found in 26 recently derived individuals. Haplotype A2 was shared by seven individuals among three introduced strains. The other six haplotypes accounted for 0. 96-1. 92% of the animals. Combined with the published sequences of European rabbits, a reduced median-joining network was constructed. The Chinese rabbit mtDNAs were scattered into two clusters of European rabbits. These results suggest that the (so-called) Chinese rabbits were introduced from Europe. Genetic diversity in Chinese rabbits was very low.

Phylogenetic relationships of Drosophila melanogaster species group deduced from spacer regions of histone gene H2A-H2B

Nucleotide sequences of the spacer region of the histone gene H2A-H2B from 36 species of Drosophila melanogaster species group were determined. The phylogenetic trees were reconstructed with maximum parsimony, maximum likelihood, and Bayesian methods by u