松辽黑猪NR5A2基因多态性及其与繁殖性状的关联分析

试验旨在探究松辽黑猪NR5A2基因多态性及其与繁殖性状的关联性。选取130头松辽黑猪作为研究对象,利用PCR直接测序法查找NR5A2基因6个外显子的SNP位点,通过HRM分型技术分析SNP位点的多态性,使用SPSS 19.0软件分析NR5A2基因SNP位点多态性与母猪总产仔数、产活仔数、仔猪初生重、3周龄重、断奶重及断奶仔猪数、乳头数的关联性。结果显示,松辽黑猪NR5A2基因第6外显子上存在1个SNP位点（C99T）,检测到3种基因型：CC、CT和TT。群体遗传参数分析及卡方适合性检验结果显示,该SNP位点在松辽黑猪群体中处于Hardy-Weinberg平衡状态（P>0.05）,但C/T突变位点杂合度相对较低,在松辽黑猪群体中的变异较小,属于中度多态（0.25<PIC<0.5）。关联性分析结果表明,在NR5A2基因C99T位点上,CC基因型个体的总产仔数、产活仔数及断奶仔猪数均最高,且显著高于TT基因型（P<0.05）,产活仔数及断奶仔猪数均显著高于CT基因型（P<0.05）;各基因型仔猪初生重、3周龄重、断奶重及乳头数均无显著差异（P>0.05）。综上所述,NR5A2基因第6外显子存在突变位点C99T,与松辽黑猪产仔数存在显著关联性,但该突变位点能否作为松辽黑猪产仔数的遗传标记,需要对更大的群体进行进一步研究。     

猪miR-199a-5p对脂质生成的影响及其靶基因的预测和分析

本研究旨在分析miR-199a-5p对猪肌内脂肪细胞脂质生成的影响及作用机制。采集淮南猪不同育肥时期（育肥前期、中期和后期）的背最长肌和皮下脂肪组织,通过实时荧光定量PCR分析其表达变化趋势;合成miR-199a-5p的mimics,转染猪原代肌内脂肪细胞,诱导分化后通过油红O染色观察过表达miR-199a-5p对脂质生成的影响;结合之前筛选到的脂肪型和瘦肉型猪差异表达mRNAs、lncRNAs和circRNAs,使用miRanda软件筛选miR-199a-5p的靶分子,并用GO和KEGG富集分析法对这些mRNAs、lncRNAs的共表达基因和circRNAs的来源基因进行功能分析。结果显示,随着育肥进程,miR-199a-5p在背最长肌中表达量持续上调,而在皮下脂肪组织中表达量先下调后上调;过表达miR-199a-5p可抑制肌内脂肪细胞的脂质生成。共筛选到9个mRNAs、5个lncRNAs和1 258个circRNAs包含miR-199a-5p结合位点,GO分析主要富集于内质网和肌动蛋白结合,KEGG分析发现主要富集于糖、脂质和蛋白质代谢。提示miR-199a-5p可能通过与lncRNAs和circRNAs互作间接调控靶基因,参与调控肌肉发育、脂质生成和代谢,可作为影响肉质性状的候选miRNA。     

5-azaC对萝卜茎尖DNA甲基化和开花的影响

本文研究了去甲基化因子5氮杂胞苷(5azaC)对萝卜开花及幼苗茎尖DNA甲基化水平的影响。用0(对照)、0.10、0.25、0.50和1.00mmolL5azaC处理‘短叶13’萝卜种子6d,除对照外,茎尖组织DNA甲基化水平均有所降低,并与春化后茎尖组织DNA甲基化水平相当;茎尖组织DNA甲基化水平随着处理浓度的提高而下降。同时,5azaC处理明显促进春性品种萝卜‘短叶13’的开花。结果表明5azaC可以部分代替低温诱导萝卜开花。     

Variation and phylogeny of the ribosomal DNA unit types and 5 S DNA inPetunia Jussieu

Summary Seven wildPetunia species including 2n = 18 species (P. parviflora Jussieu,P. linearis Hook.) and those with 2n = 14 (P. parodii Steene,P. axillaris Lam.,P. integrifolia Hook.,P. inflata R.E. Fries,P. violacea Lindl.) and tenPetunia hybrida horticultural lines were compared for polymorphisms in rDNA genes using the four restriction enzymesEcoRI,BamHI,HindIII andXhoI. All the unit types found in the lines pre-existed in the wild forms. There are two different sizes of either 11.45 or 11.6 kb./The 2n = 18 species are closely related to the 2n = 14 species, thus making thePetunia genus homogeneous. Moreover, it is likely thatP. hybrida lines originated in several kinds of crosses between these species. We constructed a dendrogram for all the 15 rDNA unit types found. Two main branches of the tree result from the presence or the absence ofHindIII sites. The main branch is divided according to variability at theEcoRI andBamHI sites. Taking into account the existence of several loci which carry one unit type only, we consider whether or not exchanges might occur between loci. Lines carrying two unit types and lines carrying three unit types support such a hypothesis.XhoI andBamHI fragments enable us to distinguish two types of 5S DNA corresponding to 2n = 18 and 2n = 14 species, respectively.P. hybrida lines and each 2n = 14 wild species carry one of the types only, that corresponds to one 5S DNA locus. The most parsimonious phylogenetic trees whatever the species chosen as the outgroup, do not fit with our knowledge ofPetunia and with taxonomy. This is likely because only few loci formed the basis of these phylogenetic constructions.     

H5N1型禽流感病毒血凝素HA1蛋白在苏云金芽胞杆菌细胞表面的展示及其免疫原性

利用苏云金芽胞杆菌S-层蛋白CTC表面展示系统研究在细胞表面展示H5N1型禽流感病毒HA1蛋白的可行性及其免疫原性，为研制既安全有效又能常温长期保藏和运输的禽流感口服基因工程疫苗奠定基础。用部分ha1基因（ha1p）代替S-层蛋白ctc基因中部且位于表面锚定序列slh下游的片段，构建了融合基因ctc-ha1p和 csa-ctc-ha1p；利用电脉冲转化法将含融合基因的重组质粒转入苏云金芽胞杆菌无质粒突变株BMB171中，获得了重组菌株BCCH（含ctc-ha1p，并导入一个载有协助细胞表面展示的csaAB操纵子的质粒）和CH（含csa-ctc-ha1p）。通过血凝和血凝抑制试验以及小鼠免疫学实验证实，2个重组菌株均成功地在细胞表面展示了重组HA1蛋白并具有一定的特异性和免疫原性，其中，CH的效果强于BCCH，说明csa-ctc-ha1p这种融合基因的构建方式更胜一筹。研究结果表明，苏云金芽胞杆菌S-层蛋白CTC表面展示系统可用来研制禽流感口服疫苗。     

The 5S rRNA gene sequence variation in wheats and some polyploid wheat progenitors (Poaceae: Triticeae)

We have cloned and sequenced 115 repeat units of the 5S rDNA genes and spacers from wheat (Triticum) and the polyploid wheat progenitor, Aegilops, and analyzed them together with sequences available in GenBank® (National Center for Biotechnology Information, NCBI, NLM, NIH, Bethesda, Maryland, USA). We were able to assort the sequences into nine orthologous groups which we call unit classes. The following unit classes were assigned to haplomes, and labeled accordingly: long A1, short A1, short A2, long G1, short G1, long D1, short D1, long S1 and short S1. The AA-genome, DD-genome and SS-genome species were each found to contain a long and a short class. The AAGG-genome species T. timopheevii and the AAA^tA^tGG-genome species T. zhukovskyi, both contain the long A1, long G1 and short G1 unit classes. The AABB-genome species T. turgidum consists of the short A2, a unit class not yet found in T. monococcum, and the long S1 unit class found in the species of Aegilops section Sitopsis. The bread wheat AABBDD-genome contains the long A1, short A2, long D1, long S1 and short G1 unit classes. The presence of the long S1, also demonstrated to occur in both T. turgidum and T. aestivum, supports the hypothesis that the progenitor of the B-haplome in wheat originated in Aegilops section Sitopsis. The presence of the short G1 unit class, i.e. the G-haplome in bread wheat, is unexpected. These new findings are discussed in the light of published findings, especially those relating to 5S DNA loci and evolutionary hypotheses.     

Biodiversity of micro-eukaryotes in Antarctic Dry Valley soils with <5% soil moisture

Soils in the Dry Valleys of Antarctica are considered to be among the world's most extreme environments. These soils are old, cold and dry with low contents of organic carbon and nitrogen. Habitats adjacent to water (lakes and ice melts) have significant biological activity as demonstrated by the presence of algal mats, lichens and small invertebrates, particularly nematodes, tardigrades and rotifers. In contrast, there are extensive areas in the Dry Valleys that are extremely dry with less than 5% moisture content. These soils are often salty and appear to be barren of life as they have a coarse texture due to their lack of plant organic material. In contrast, molecular techniques (DNA extraction from soils, cloning and rDNA sequence analysis) demonstrated the presence of a complex micro-eukaryotic food web whose structure and composition varied with moisture content and location. Micro-eukaryotic communities in soils with 0.2-1.3% moisture were represented by species of the yeast genus Trichosporon and an unidentified clade of micro-eukaryotes, whereas levels from 3.1% to 4.9% contained complex food webs including primary producers (chlorophytes and stramenopiles), symbionts (lichen associated fungi), saprophytes (fungi), predators (alveolates and cercozoans) and fungal nematode parasite/pathogens. The soils had a diversity of species (80 species from 15 sites) with a restricted number (3-21 species) at each site. The sensitive and measurable community structure of the low moisture Dry Valley soils provides an unparalleled opportunity to examine local and global environmental effects on micro-eukaryotic community dynamics with multiple trophic levels.     

小麦体细胞组织离体诱变效应研究

小麦种子经30kRγ射线照射后,再从M1株取未成熟胚培养,用再生株当代(M2R1)的死苗率、不育株率、结实率及株高变化所反映的损伤程度与对照株（R1）比较，无明显差异；M3R2代的变异率也与对照R2相仿。5天龄幼胚用0.5kR照射处理后的再生株M1R1的损伤程度与对照相仿；M2R2的变异率随不同基因型而有增减，但变异率仍有显著提高。未     

Biolog法测定土壤微生物群落功能多样性预处理方法的筛选

作为测定微生物群落功能多样性主要方法之一的biolog法,由于操作简单、灵敏度高、分辨力强、获得数据量丰富,受到众多研究者的青睐,但文献中报道的有关实验预处理方法却不尽相同。为了筛选最佳的预处理方法,通过比较文献中3种较为常用的方法,研究不同预处理对biolog法测定微生物群落功能多样性的影响。结果表明:AWCD值总体变化趋势B方法优于A、C;微生物对六类碳源的利用率中B方法对大部分碳源利用较高,且结果较稳定;两种处理的Simpson指数、Shannon丰富度指数及碳源利用丰富度指数均BCA,而Mc Intosh指数、Shannon均匀度结果不太稳定,但经方差分析,其差异不显著,且B方法的方差最小;主成分分析的结果 B方法优于A与C。总之,在250 r/min的转速下震荡30 min,并于28℃条件下培养接种平板的处理(B法)效果较好,其测定结果重现性好,且操作简便快捷,是比较理想的预处理方法。