构树种质资源遗传多样性SRAP分子标记研究

构树（Broussonetia papyrifera L.）为桑科（Moraceae）构树属（Broussonetia）落叶乔木，广泛分布于亚洲东部及太平洋岛屿。它是我国重要的经济林木，具有重要的经济价值，其树皮纤维品质优良，自古就是造纸的优良原料；叶片可用作饲料；果实具有重要的药用价值；环境适应性强，是迅速绿化荒山、荒滩和盐碱地的理想树种。因此对构树这些特性的深入研究和开发利用具有非常重要的实际应用价值。本研究以日本和国内构树主要分布区域的的10种生态型及杂交构树共23份材料，摸索并改进了构树DNA的提取方法，建立了稳定的SRAP分子标记体系，以杂交构树组培苗为材料从120对引物组合中筛选出条带信息较高的17对SRAP引物，以这些引物对23份构树材料进行PCR扩增和标记分析。 本研究取得的主要结果如下： （1）本实验首次将SRAP技术应用于构树的研究中，建立起构树稳定的SRAP-PCR反应体系；实验中对影响扩增的5个主要因素进行了优化，确定20 μL PCR反应体系中各因素最适浓度：模板DNA浓度80 ng/(20 μL)，Mg2+浓度2.0 mM，dNTP浓度0.6 mM，引物浓度0.8 mM，Taq 酶浓度1.5 U/(20 μL)。 （2）从120对引物组合中筛选出来的17对SRAP引物对21份不同生态型构树样本（21份材料指的是除两份杂交构树材料外的其它生态型构树，以下同）进行PCR扩增，共扩增出439条带，平均每对引物25.5条，其大小介于100～1,000 bp之间，其中多态性条带319条，占总数的72.67%。 （3）用Popgene1.32软件进行分析，计算出Shannon信息指数（I）值为0.2275（0.2042），物种水平的Nei基因多样性（H）值为0.1336（0.1436），表明各生态型构树之间的平均遗传多态性不高，中国大陆各生态型构树Shannon信息指数（I）值仅为0.1675（0.2271），物种水平的Nei基因多样性（H）值为0.1039（0.1540），群体遗传多样性较低，构树的遗传分化主要存在于中国和日本之间。 （4）用NTSYS-2.10e软件进行聚类分析，发现不同生态型构树按距离关系远近及分布区域可划分为不同类群。在遗传相似系数0.57附近，对21份材料进行聚类分析发现其可分为两个群体，一类为日本生态型，另一类为中国生态型，表明日本构树与中国野生种构树种源遗传相似性较小。中国各生态型构树在遗传相似系数0.91处可分为5类，总体而言，中国各地区之间的构树遗传相似度较高。对杂交构树分析表明，其亲缘关系与日本构树（母本）更接近。 （5）日本及杂交构树的SCAR标记。本研究找到两条日本及杂交构树的特异性条带，回收、测序，再根据序列往里重新设计引物，转变成稳定性更好，更直观的SCAR标记，这为挑选性状优良的日本及杂交构树提供重要的参考，对其育种有一定的指导意义。其中一条片段经与NCBI数据库比对发现与拟南芥磺基转移酶家族基因具有较高的同源性。

PLOD3基因3'调控区的人类特异突变改变miR-124a对PLOD3的调控

microRNAs(miRNAs)是一类在转录后水平调控基因表达的不编码蛋白质的小RNA(长度20-24个碱基).其中,miR-124a是一个在哺乳动物中枢神经系统高度表达的miRNA,在神经前体细胞向神经元分化的过程中起着举足轻重的作用.由于miRNAs特异性地识别靶基因的3'端调控区(3'UTR)的靶序列,因此,在人类起源过程中基因3'UTR的单核苷酸序列变异有可能导致miRNA调控的改变.通过靶基因预测和3'UTR区在哺乳动物代表物种间的同源序列比较,我们发现miR-124a的靶基冈中有一个基因(PLOD3)3UTR的靶位点中存在人类特异突变位点.利用体外报告基因系统,发现PLOD3基因3'UTR靶位点中所含的一个人类特异的突变导致miR-124a对PLOD3的调控效率降低.研究表明,miRNAs靶基因3'UTR的序列变异具有功能效应,它有可能足人类中枢神经系统在起源和演化中发挥关键作用的重要遗传机制之一.

The mitochondrial genome organization of the rice frog, Fejervarya limnocharis (Amphibia : Anura): a new gene order in the vertebrate mtDNA

The mitochondrial DNA of the rice frog, Fejervarya limnocharis (Amphibia, Anura), was obtained using long-and-accurate polymerase chain reaction (LA-PCR) combining with subcloning method. The complete nucleotide sequence (17,717 bp) of mitochondrial genome was determined subsequently. This mitochondrial genome is characterized by four distinctive features: the translocation of ND5 gene, a cluster of rearranged tRNA genes (tRNA(Thr), tRNA(Pro), tRNA(Leu) ((CUN))) a tandem duplication of tRNA(Mer) gene, and eight large 89-bp tandem repeats in the control region, as well as three short noncoding regions containing two repeated motifs existing in the gene cluster of ND5/tRNA(Thr)/tRNA(Pro)/tRNA(Leu)/tRNA(Phe). The tandem duplication of gene regions followed by deletions of supernumerary genes can be invoked to explain the shuffling of tRNAM(Met) and a cluster of tRNA and ND5 genes, as observed in this study. Both ND5 gene translocation and tandem duplication of tRNA(Met) were first observed in the vertebrate mitochondrial genomes. (c) 2004 Elsevier B.V. All rights reserved.

17 beta-Estradiol and progesterone improve in-vitro cytoplasmic maturation of oocytes from unstimulated prepubertal and adult rhesus monkeys

BACKGROUND: Effects of 17beta-estradiol and progesterone on rhesus monkey oocyte maturation in vitro were evaluated by embryo development subsequent to IVF. METHODS AND RESULTS: In experiment 1, immature cumulus-oocyte complexes collected from unstimulated adult females during the non-breeding season were matured in modified medium CMRL-1066 containing various combinations of gonadotrophins (FSH + LH), estradiol and/or progesterone. Formation of morulae and blastocysts was greatest in oocytes matured in medium containing estradiol and/or progesterone, with or without gonadotrophins (morula 38-46%, blastocyst 14-20%) than in control oocytes matured without estradiol or progesterone (morula 14%, blastocyst 0%). In experiment 2, cumulus-oocyte complexes from unstimulated prepubertal female monkeys were matured in medium with gonadotrophins, estradiol or progesterone. The best development to the morula stage was obtained with oocytes matured with gonadotrophins and estradiol or gonadotrophins and progesterone (43 and 25 morulae, respectively), while control oocytes matured with gonadotrophins but without steroid hormones gave the poorest morula developmental response (12%). However, there was no difference in blastocyst development across all groups (0-3%). CONCLUSIONS: These results demonstrate that during rhesus monkey oocyte maturation in vitro: (i) estradiol or progesterone can improve oocyte developmental competence; (ii) immature oocytes from prepubertal versus adult females have differential responses to challenge with estradiol or progesterone.

Bacillus pallidus sp nov., isolated from forest soil

A Gram-positive bacterium, designated strain CW 7(T), was isolated from forest soil in Anhui Province, south-east China. Cells were strictly aerobic, motile with peritrichous flagella and rod-shaped. The strain grew optimally at 30-37 degrees C and pH 7.0-8.0. The major fatty acids of strain CW 7(T) were anteiso-C-15:0, iso-C-15:0 and anteiso-C-17:0. The predominant menaquinone was MK-7. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The G + C content of the genomic DNA was 42.3 mol%. Phylogenetic analysis indicated that strain CW 7(T) belonged to a monophyletic cluster within the genus Bacillus and showed 16S rRNA gene sequence similarities of less than 96.5% to recognized species of the genus Bacillus. The results of the polyphasic taxonomic study, including phenotypic, chemotaxonomic and phylogenetic analyses, showed that strain CW 7(T) represents a novel species of the genus Bacillus, for which the name Bacillus pallidus sp. nov. is proposed. The type strain is CW 7(T) (=KCTC 13200(T)=CCTCC AB 207188(T)=LMG 24451(T)).

Degradation of tropical forest in Hainan, China, 1991-2008: Conservation implications for Hainan Gibbon (Nomascus hainanus)

The Hainan gibbon (Nomascus hainanus) is one of the most endangered primates in the world, confined to mature natural forest in Hainan Island, China. We assessed changes in habitat condition on the island between 1991 and 2008, using vegetation maps generated by remote-sensing images. We defined forest suitable for gibbons based on composition, tree size and canopy cover. During the 17-year period, the area of suitable gibbon forest decreased by 540 km(2) (35%) across the whole island, and by 6.3 km(2) (7%) in the locality of the sole remaining gibbon population at Bawangling National Nature Reserve. The forest patches large enough (>1 km(2)) to support a gibbon group decreased from 754 km(2) to 316 km(2) in total area, and from 92 to 64 in number. Suitable natural forest was mainly replaced by plantations below 760 m, or degraded by logging, grazing and planting of pines above 760 m. Meanwhile, forests in former confirmed gibbon areas became more fragmented: mean area of patches decreased by 53%. We mapped the patches of natural forest in good condition which could potentially support gibbons. We recommend a freeze on further expansion of plantations between core patches at Bawangling, Jiaxi-Houmiling and Yinggeling Nature Reserves in accordance with forest protection regulations; establishment of nature reserves in currently unprotected natural forest patches elsewhere in line with the local government's nature reserve expansion policy; and active natural-forest restoration between remaining fragments at Bawangling. (C) 2010 Elsevier Ltd. All rights reserved.

Polymorphisms in the C-type lectin genes cluster in chromosome 19 and predisposition to severe acute respiratory syndrome coronavirus (SARS-CoV) infection

Background: Polymorphisms of CLEC4M have been associated with predisposition for infection by the severe acute respiratory syndrome coronavirus (SARS-CoV). DC-SIGNR, a C-type lectin encoded by CLEC4M, is a receptor for the virus. A variable number tandem

17种植物中蛋白质提取物的抗HIV-1活性

以化合物对ＨＩＶ１诱导Ｃ８１６６细胞形成合胞体的抑制实验和化合物对ＨＩＶ１感染细胞的保护实验作为初筛方法，筛选了来源于７科１７种植物的４７个样品，其中８个样品经测定是核糖体失活蛋白（ＲＩＰｓ），其余为粗提蛋白。进而测定了初筛有抗ＨＩＶ活性的化合物对共培养、急性和慢性感染的ＨＩＶ１ｐ２４抗原表达水平的影响，用间接荧光染色检测这些化合物对ＨＩＶ抗原阳性细胞率的影响，以确证其抗ＨＩＶ活性。天花粉蛋白（ＴＣＳ）、南方栝楼蛋白峰Ⅴ、南方栝楼蛋白峰Ⅵ均显著地抑制合胞体的形成；巴Ⅱ、丝瓜子蛋白、油瓜根蛋白、木盍藤子蛋白有一定的抑制作用。老鼠拖瓜蛋白、大叶木鳖子根蛋白、西双版纳根Ⅱ等粗提蛋白有显著的抑制作用。ＲＩＰｓ不能保护ＨＩＶ１感染细胞的死亡。ＴＣＳ显著地抑制了ＨＩＶ１急性感染中ｐ２４抗原的表达，减少了ＨＩＶ抗原阳性细胞数，但它们均不影响共培养细胞融合、ＨＩＶ１慢性感染中ｐ２４抗原表达水平。结果表明，除ＴＣＳ等已知ＲＩＰｓ外，还有一些新的ＲＩＰｓ具有不同程度的抗ＨＩＶ活性。

Molecular phylogeny and taxonomy of wood mice (genus Apodemus Kaup, 1829) based on complete mtDNA cytochrome b sequences, with emphasis on Chinese species

Phylogenetic relationships among 15 species of wood mice (genus Apodemus) were reconstructed to explore some long-standing taxonomic problems. The results provided support for the monophyly of the genus Apodemus, but could not reject the hypothesis of paraphyly for this genus. Our data divided the 15 species into four major groups: (1) the Sylvaemus group (A. sylvaticus, A. flavicollis, A. alpicola, and A. uralensis), (2) the Apodemus group (A. peninsulae, A. chevreri, A. agrarius, A. speciosus, A. draco, A. ilex, A. semotus, A. latronum, and A. mystacinus), (3) A. argenteus, and (4) A. gurkha. Our results also suggested that orestes should be a valid subspecies of A. draco rather than an independent species; in contrast, A. ilex from Yunnan may be regarded as a separate species rather than a synonym of orestes or draco. The species level status of A. latronum, tscherga as synonyms of A. uralensis, and A. chevrieri as a valid species and the closest sibling species of A. agrarius were further corroborated by our data. Applying a molecular clock with the divergences of Mus and Rattus set at 12 million years ago (Mya) as a calibration point, it was estimated that five old lineages (A. mystacinus and four major groups above) diverged in the late Miocene (7.82-12.74 Mya). Then the Apodemus group (excluding A. mystacinus) split into two subgroups: agrarius and draco, at about 7.17-9.95 Mya. Four species of the Sylvaemus group were estimated to diverge at about 2.92-5.21 Mya. The Hengduan Mountains Region was hypothesized to have played important roles in Apodemus evolutionary histories since the Pleistocene. (C) 2004 Elsevier Inc. All rights reserved.

大熊猫生殖生物学研究 I:大熊猫发情期血清和尿液中促黄体素、孕酮和17β-二雌二醇含量的变化

放射免疫和生物测定法表明3只大熊猫在发情高峰期均出现雌二醇和促黄体素高峰; LH峰值出现在E_(2)峰值以后。根据雌兽在发情期血清和尿液中这二种激素的变化可为选择人工授精最佳时间提供可靠的理论依据。图2表2参15

鸟类核型研究Ⅸ. 画眉亚科17种

该文报道了雀形目鶲科画眉亚科17种鸟类核型, 臂间倒位和小染色体缺失在该亚科进化中起重要作用. 在核型比较基础上结合古地理以及生态资料对该类群的起源和进化作了初步探讨。

利用17个微卫星标记分析鳙鱼的遗传多样性

选用实验室克隆的17个鳙鱼微卫星分子标记分析四川泸州和江西鄱阳湖的两个种群鳙鱼的遗传多样性及种质特性,计算和统计了杂合度、多态信息含量(PIC)、有效等位基因数、等位基因频率、遗传距离、遗传相似系数、Hardy-Weinberg平衡偏离指数等方面内容。结果表明:选择使用17个微卫星标记,其中有4个为单态标记,13个为多态标记。江西和四川鳙鱼群体每个微卫星位点的平均等位基因数分别为3.325及3.882,平均有效等位基因数分别为3.531及2.676,多态位点百分率分别为82.4及70.5,17个微卫星标记

黑尾近红鲌含肉率及肌肉营养成分分析

测定了长江上游特有鱼类黑尾近红的含肉率及其肌肉生化成分和能值。结果表明 :黑尾近红的含肉率为 70 74 % ;其肌肉生化成分 (鲜重百分比 )水分含量为 80 97%、蛋白质 15 92 %、脂肪 1 11%、灰分 1 0 1%、无氮浸出物 0 99%、比能值 4 37kJ/g及E/P 2 7 4 5kJ/g。肌肉生化成分含量与体长变化无明显关系。肌肉中含有 17种氨基酸 ,总量为 6 9 0 7% (干重百分比 ) ;9种人体必需氨基酸总量是 34 81% ,必需氨基酸占氨基酸总量比例为5 0 3