冬小麦不同生育时期水分亏缺胁迫对叶片保护酶系统的影响

利用盆栽试验研究了施Ｎ（底肥）与不施Ｎ条件下冬小麦不同生育时期水分有限亏缺对叶片保护酶系统的影响,结果表明,在无底肥条件下,３个生育时期水分亏缺均使保护酶系统活性有所降低,而在有底肥时酶活性多数则升高,各保护酶活性与ＭＤＡ相关分析表明,苗期ＳＯＤ活性与ＭＤＡ呈极显著负相关;拔节却是ＰＯＤ活性与ＭＤＡ呈极显著负相关;在灌浆期ＳＯＤ、ＰＯＤ、ＣＡＴ与ＭＤＡ含量均无显著相关性,但ＭＤＡ在叶片中累积至较高水平。各生育期水分处理叶片绿素含量与ＭＤＡ含量达极显著负相关。这些说明在不同生育时期,受水分亏缺和供的,保护酶系统各酶的变化有明显差异。其生理作用也有差异。     

Improving thermal hysteresis activity of antifreeze protein from recombinant Pichia pastoris by removal of N-glycosylation

To survive in a subzero environment, polar organisms produce ice-binding proteins (IBPs). These IBPs prevent the formation of large intracellular ice crystals, which may be fatal to the organism. Recently, a recombinant FfIBP (an IBP from Flavobacterium frigoris PS1) was cloned and produced in Pichia pastoris using fed-batch fermentation with methanol feeding. In this study, we demonstrate that FfIBP produced by P. pastoris has a glycosylation site, which diminishes the thermal hysteresis activity of FfIBP. The FfIBP expressed by P. pastoris exhibited a doublet on SDS-PAGE. The results of a glycosidase reaction suggested that FfIBP possesses complex N-linked oligosaccharides. These results indicate that the residues of the glycosylated site could disturb the binding of FfIBP to ice molecules. The findings of this study could be utilized to produce highly active antifreeze proteins on a large scale.     

基因组重排技术选育乙醇高产菌株

里氏木霉（Trichoderma reesei）被认为是最合适联合生物加工（consolidated bioprocessing）的微生物之一。原始里氏木霉菌株产乙醇能力太低,需要进一步提高其产酒量。我们通过基因组重排技术提高了里氏木霉菌株产乙醇能力和乙醇耐受力。首先对CICC40360菌株孢子进行NTG诱变得到正向突变菌株,再以此为出发菌株进行基因组重排。进行基因组重排后,重组菌株在含不同乙醇浓度的原生质体再生培养基上进行筛选。突变菌株和原始菌株一起做摇瓶发酵实验进行比较以确定产乙醇能力的提高。经过两轮基因组重排后,筛选获得表现最优异的重组菌S2-254。该菌株能在利用50g/l葡萄糖发酵出6.2g/l乙醇,同时能耐受3.5% （v/v）浓度乙醇。上述结果表明,本实验采用的基因组重排技术能够有效而且快速获得具有目的性状的优良菌株。     

木本植物基因组DNA提取及鉴定

采用改良后的CTAB法,对山葡萄、软枣猕猴桃、蒙古栎、核桃楸、西伯利亚红松和偃松基因组DNA进行提取。结果表明,所提基因组DNA分子量与λDNA（48 kb）接近,其紫外吸收比在1.66～1.89之间。第3次和第4次上清提取的DNA质量优于第1次和第2次。从提取产量看,每克鲜重提取DNA量最小为15 μg·g^-1（核桃楸第4次上清）,最高的为272 μg·g^-1（山葡萄第3次上清）。西伯利亚红松和偃松第1次和第2次上清基本未提出DNA,第3次和第4次上清中得到了较高质量的DNA。经酶切鉴定和PCR扩增,所提的基因组DNA可以用于进一步研究。     

Seroprevalence of Toxoplasma gondii in wild boars (Sus scrofa leucomystax) and wild sika deer (Cervus nippon) in Gunma Prefecture, Japan

The ingestion of undercooked meat from wild animals can be a source of Toxoplasma gondii infection in humans and other animals. In this study, we determined the seroprevalence of T. gondii infection in 175 wild boars (Sus scrofa leucomystax) and 107 wild sika deer (Cervus nippon) hunted in 2004–2007 in Gunma Prefecture, Japan, by using a commercial latex agglutination test (LAT). Antibodies (LAT, 1:64 or higher) to T. gondii were found in 6.3% of wild boars and 1.9% of sika deer. This is the first record of T. gondii infection in wild deer in Japan, and deer and wild boar meat should be cooked well before human consumption.     

An Efficient reproductive method for Irs2-/- mice with C57BL/6JJcl genetic background

Efficient reproduction using natural mating and reproduction technology [in vitro fertilization (IVF) and embryo transfer (ET)] was investigated in IRS2 deficient mice with C57BL/6JJcl genetic background (Irs2(-/-) mice) as a typical type 2 diabetes model. From the results using various combinations of Irs2(-/-) and Irs2(-/+) mice, the combination of female Irs2(-/+) x male Irs2(-/-) was found to be more efficient than other combinations. In applications of reproduction technology using IVF and ET, the combination of female Irs2(-/+) x male Irs2(-/-) involves the possibility of Irs2(-/-) production by repeats using female Irs2(-/+) mice. However, reproductive continuity using this combination is difficult because of dependence on human technique and the cost of ET. Therefore, we concluded that Irs2(-/-) mice should be produced by embryo transfer using Irs2(-/-) mice from a colony consisting of female Irs2(-/+) x male Irs2(-/-).     

Structure and dynamics of small soluble Aβ(1-40) oligomers studied by top-down hydrogen exchange mass spectrometry

Aβ peptides can assemble into amyloid fibrils, which represent one of the hallmarks of Alzheimer's disease. Recent studies, however, have focused on the behavior of small soluble Aβ oligomers that possess a much greater neurotoxicity than mature fibrils. The structural characterization of these oligomers remains difficult because of their highly dynamic and polymorphic nature. This work explores the behavior of Aβ(1-40) in a slightly basic solution (pH 9.3) at a low salt concentration (10 mM ammonium acetate). These conditions lead to the formation of small oligomers, without any signs of fibrillation for several hours. The structure and dynamics of these oligomers were characterized by circular dichroism spectroscopy, size exclusion chromatography, and millisecond time-resolved hydrogen exchange mass spectrometry (MS). Our results reveal rapid interconversion between Aβ(1-40) oligomers and monomers. The mole fraction of monomeric molecules is on the order of 40%. Oligomers consist of ~4 Aβ(1-40) molecules on average, and the resulting assemblies have a predominantly β-sheet secondary structure. Hydrogen exchange proceeds in the EX1 regime. This feature allows the application of conformer-specific top-down MS. Electron capture dissociation is used for interrogating the deuteration behavior of the Aβ(1-40) oligomers. This approach provides a spatial resolution of ~2 residues. The backbone amide deuteration pattern uncovered in this way is consistent with a β-turn-β motif for L17-M35. The N-terminus is involved in hydrogen bonding, as well, whereas protection gradually tapers off for C-terminal residues 35-40. Our data are consistent with earlier proposals, according to which Aβ(1-40) oligomers adopt a β-barrel structure. In general terms, this study demonstrates how top-down MS with precursor ion selection can be employed for structural studies of specific protein conformers within a heterogeneous mix.     

Rotary torque produced by proton motive force in FoF1 motor

We have attempted direct observation of the light-driven rotation of a FoF(1)-ATP motor. The FoF(1)-ATP motor was co-reconstituted by the deletion-delta subunit of FoF(1)-ATP synthase with bacteriorhodopsins (BRs) into a liposome. The BR converts radiation energy into electrochemical gradient of proton to drive the FoF(1)-ATP motor. Therefore, the light-driven rotation of FoF(1)-ATP motor has been directly observed by a fluorescence microscopy using a fluorescent actin filament connected to beta-subunit as a marker of its orientation. The rotational torque value of the Fo motor was calculated as 27.93+/-1.88pNnm. The ATP motor is expected to be a promising rotary molecular motor in the development of nanodevices.     

西南岩溶地区黄荆和檵木叶片结构对其生态环境的响应

应用常规石蜡切片法对生长于桂林毛村岩溶区和非岩溶区的黄荆(Vitex negundo)和檵木(Loropetalumchinense)的解剖特征进行了比较研究,并对两区的黄荆叶片表皮形态进行了扫描电镜观察.结果显示:(1)两地的黄荆叶片背面均有浓密的绒毛,但致密程度有差异,岩溶区黄荆叶片的气孔深藏于绒毛间隙,这种结构可减少水分蒸发,降低因岩溶干旱带来的水分缺失.(2)岩溶区黄荆和檵木的叶片厚度、上下表皮厚度、栅栏组织的厚度以及栅栏组织的致密程度均大于非岩溶区,这些特征有利于减少水分蒸腾.(3)岩溶区黄荆和檵木叶片的维管组织发达程度高于非岩溶区,有利于在蒸腾减小的情况下促进水分运输和营养元素的迁移,说明2种植物叶片结构特征在不同生境区的改变是其长期在岩溶区干旱环境条件下形成的适应性变化.