甘薯遗传作图策略研究与展望

甘薯分子遗传图谱的建立对甘薯分子育种技术体系的拓展和应用具有重要意义。当前,对甘薯分子遗传图谱的研究虽然取得一定的进展,但存在着很多技术瓶颈,如作图策略应用和优化等。总结了甘薯经典和分子遗传研究进展,剖析了甘薯分子遗传图谱作图的3种方法与策略;探讨和提出了提高甘薯作图效率和质量的途径主要是:优化作图群体质量、克服偏分离、整合多群体间遗传连锁图谱和选择合适的分子标记类型;并指出染色体关联在遗传作图中的重要性,提出甘薯分子育种领域亟待加强的方面,以期为今后甘薯精密分子图谱的建立及基于分子图谱的甘薯分子育种提供新的思路。     

Analysis of variable sites between two complete South China tiger (Panthera tigris amoyensis) mitochondrial genomes

In order to investigate the mitochondrial genome of Panthera tigris amoyensis, two South China tigers (P25 and P27) were analyzed following 15 cymt-specific primer sets. The entire mtDNA sequence was found to be 16,957 bp and 17,001 bp long for P25 and P27 respectively, and this difference in length between P25 and P27 occurred in the number of tandem repeats in the RS-3 segment of the control region. The structural characteristics of complete P. t. amoyensis mitochondrial genomes were also highly similar to those of P. uncia. Additionally, the rate of point mutation was only 0.3% and a total of 59 variable sites between P25 and P27 were found. Out of the 59 variable sites, 6 were located in 6 different tRNA genes, 6 in the 2 rRNA genes, 7 in non-coding regions (one located between tRNA-Asn and tRNA-Tyr and six in the D-loop), and 40 in 10 protein-coding genes. COI held the largest amount of variable sites (9 sites) and Cytb contained the highest variable rate (0.7%) in the complete sequences. Moreover, out of the 40 variable sites located in 10 protein-coding genes, 12 sites were nonsynonymous.     

Aptamer selection for the detection of Escherichia coli K88

In this study, the first group of single-stranded DNA aptamers that are highly specific to enterotoxigenic Escherichia coli (ETEC) K88 was obtained from an enriched oligonucleotide pool by the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) procedure, during which the K88 fimbriae protein was used as the target and bovine serum albumin as counter targets. These aptamers were applied successfully in the detection of ETEC K88. They were then grouped under different families based on the similarity of their secondary structure and the homology of their primary sequence. Four sequences from different families were deliberately chosen for further characterization by fluorescence analysis. Having the advantage of high sensitivity, fluorescence photometry was selected as single-stranded DNA quantification method during the SELEX process. Aptamers with the highest specificity and affinity were analyzed to evaluate binding ability with E. coli. Since ETEC K88 is the only type of bacterium that expressed abundant K88 fimbriae, the selected aptamers against the K88 fimbriae protein were able to specifically identify ETEC K88 among other bacteria. This method of detecting ETEC K88 by aptamers can also be applied to bacteria other than ETEC K88.     

Human liver alkaline phosphatase, purification and partial sequencing: homology with the placental isozyme

Human liver alkaline phosphatase (AP) has been purified to homogeneity. The enzyme has a molecular weight of 150,000 in its native state and consists of two identical subunits of Mr 75,000. After treatment with endoglycosidase F the molecular weight is reduced to 50,000 indicating a high degree of glycosylation. The amino-terminal sequence up to 22 residues was found to be Leu-Val-Pro-Glu-Lys-Glu-Lys-Asp-Pro-Lys-Tyr-(Ala)-Arg-Asp-Gln-Ala-Gln-?- Thr-Leu-Lys-Tyr. The amino-terminal portions of human and bovine liver AP are identical. The amino termini of the human liver and human placental AP isozymes have appreciable homology. Conformationally the amino termini are very similar.     

Coating and selective deposition of nanofilm on silicone rubber for cell adhesion and growth

A recently developed method for surface modification, layer-by-layer (LbL) assembly, has been applied to silicone, and its ability to encourage endothelial cell growth and control cell growth patterns has been examined. The surfaces studied consisted of a precursor, with alternating cationic polyethyleneimine (PEI) and anionic sodium polystyrene sulfonate (PSS) layers followed by alternating gelatin and poly-d-lysine (PDL) layers. Film growth increased linearly with the number of layers. Each PSS/PEI bilayer was 3 nm thick, and each gelatin/PDL bilayer was 5 nm thick. All layers were more hydrophilic than the unmodified silicone rubber surface, as determined from contact angle measurements. The contact angle was primarily dictated by the outermost layer. Of the coatings studied, gelatin was the most hydrophilic. A film of (PSS/PEI)₄/(gelatin/PDL)₄/ gelatin was highly favorable for cell adhesion and growth, in contrast to films of (PSS/PEI)₈ or (PSS/PEI)₈/PSS. Cell growth patterns were successfully controlled by selective deposition of microspheres on silicone rubber, using microcontact printing with a silicone stamp. Cell adhesion was confined to the region of microsphere deposition. These results demonstrate that the LbL self-assembly technique provides a general approach to coat and selectively deposit films with nanometer thickness on silicone rubber. Furthermore, they show that this method is a viable technique for controlling cellular adhesion and growth.     

绵羊肌肉中FAS基因和HSL基因的发育性变化及其对肌内脂肪含量的影响

选取不同日龄的雄性哈萨克羊和新疆细毛羊共54只,屠宰后取背最长肌,用索氏抽提法检测肌内脂肪(intramuscular fat,IMF)含量,用荧光实时定量PCR法检测肌肉脂肪酸合成酶(fatty acid synthase,FAS)和激素敏感脂肪酶(hormone-sensitive lipase,HSL)基因表达的发育性变化,并分析基因表达对肌内脂肪沉积的影响。结果表明:1)随着日龄的增加,雄性哈萨克羊的IMF含量持续上升,各生长时期差异显著(P<0.05),而新疆细毛羊的IMF含量在各生长时期无显著差异(P>0.05)。雄性哈萨克羊的IMF含量30~90日龄期间极显著高于新疆细毛羊(P<0.01)。2)FAS基因mRNA水平在哈萨克羊肌肉中初生时最高(P<0.05),然后随日龄的增加呈下降趋势;在新疆细毛羊肌肉中,FAS mRNA水平表现出"下降-上升-下降-上升"的发育模式,其中60日龄显著高于90日龄(P<0.05),其余日龄之间差异不显著。HSL基因在2品种绵羊肌肉中的表达模式基本类似,在哈萨克羊肌肉中随年龄的增加而下降,初生时的水平显著高于60~90日龄(P<0.05);在新疆细毛羊中30日龄时达到最高(P<0.01),到60日龄时下降到最低(P<0.05),随后保持这种低表达水平。3)FAS和HSL基因mRNA的表达量均与哈萨克羊IMF含量呈负相关,相关系数分别为:r=-0.485(P=0.02),r=-0.423(P=0.05);在哈萨克羊中两基因表达量水平比值(FAS:HSL)与IMF呈极显著负相关r=-0.552(P=0.01)。在新疆细毛羊中两基因的表达水平及比值均与IMF无显著相关性(P>0.05)。     

中华猕猴桃和美味猕猴桃的倍性变异及地理分布研究

根据已有的猕猴桃自然地理分布资料,通过对中华猕猴桃（Actinidia chinensis）和美味猕猴桃（A．deliciosa）野外分布居群的详细调查,利用流式细胞技术对我国西部高原台地向中东部丘陵平原过渡地带6个纯中华猕猴桃、1个纯美味猕猴桃和5个中华／美味猕猴桃同域分布居群共276个个体的倍性进行了检测。结果表明：（1）中华猕猴桃存在二倍体和四倍体,美味猕猴桃存在四倍体、五倍体和六倍体;（2）中华猕猴桃和美昧猕猴桃在经度和纬度的分布上存在显著差异（P〈0．05）,中华猕猴桃在经度上偏东分布而美味猕猴桃偏西,纬度分布上中华猕猴桃偏南而美味猕猴桃偏北;而且不同倍性小种在经、纬度分布上呈现显著差异（P〈0．05）,二倍体、四倍体、六倍体的分布在经度上依次从东到西、纬度上从南到北;（3）中华猕猴桃和美味猕猴桃不同倍性小种的海拔分布存在显著性差异（P〈0．05）,二倍体小种分布海拔最低,四倍体小种次之,六倍体小种海拔分布最高,但通过LSD分析四倍体个体和六倍体个体在海拔分布上不存在显著差异（P〉0．05）。通过对中华／美味猕猴桃这两种具有重要经济价值的果树的倍性变异及地理分布的探讨,提出了猕猴桃倍性小种分布的上述规律并给猕猴桃种质资源收集、评价、创新和可持续利用方面提供了初步的研究基础,尤其是为我国猕猴桃新品种选育提供了基础数据和科学依据。     

白芨的组培快繁(简报)

本文扼要地阐述了以白芨种子为培养材料,经筛选的培养基诱导、分化、生长,最终可获得苗质好、性状均一的白芨;为进一步研究快速繁殖白芨提供参考.     

Molecular characterization of avian-like H1N1 swine influenza a viruses isolated in Eastern China, 2011

Currently, three predominant subtypes of influenza virus are prevalent in pig populations worldwide: H1N1, H3N2, and H1N2. European avian-like H1N1 viruses, which were initially detected in European pig populations in 1979, have been circulating in pigs in eastern China since 2007. In this study, six influenza A viruses were isolated from 60 swine lung samples collected from January to April 2011 in eastern China. Based on whole genome sequencing, molecular characteristics of two isolates were determined. Phylogenetic analysis showed the eight genes of the two isolates were closely related to those of the avian-like H1N1 viruses circulating in pig populations, especially similar to those found in China. Four potential glycosylation sites were observed at positions 13, 26, 198, 277 in the HA1 proteins of the two isolates. Due to the presence of a stop codon at codon 12, the isolates contained truncated PB1-F2 proteins. In this study, the isolates contained 591Q, 627E and 701N in the polymerase subunit PB2, which had been shown to be determinants of virulence and host adaptation. The isolates also had a D rather than E at position 92 of the NS1, a marker of mammalian adaptation. Both isolates contained the GPKV motif at the PDZ ligand domain of the 3′ end of the NS1, a characteristic marker of the European avian-like swine viruses since about 1999, which is distinct from those of avian, human and classical swine viruses. The M2 proteins of the isolates have the mutation (S31N), a characteristic marker of the European avian-like swine viruses since about 1987, which may confer resistance to amantadine and rimantadine antivirals. Our findings further emphasize the importance of surveillance on the genetic diversity of influenza A viruses in pigs, and raise more concerns about the occurrence of cross-species transmission events.