冷驯化条件下中缅树鼩脂肪组织代谢产物的变化

为探究冷驯化条件下中缅树鼩(Tupaia belangeri)白色脂肪组织(WAT)和褐色脂肪组织(BAT)的差异代谢物变化,本研究采集对照组和冷驯化28天组中缅树鼩的WAT和BAT,采用非靶向代谢组液相色谱—质谱联用检测技术分析其差异代谢物含量变化.结果 表明,冷驯化组较对照组WAT中有7种差异代谢物显著上调;BAT...     

云南不同地区中缅树鼩头骨形态特征的比较

中缅树鼩为东洋界热带亚热带特有类群.本研究采用几何形态测量法对分布于滇中高原(禄劝县)和横断山地区(剑川县、丽江市和云龙县)的中缅树鼩头骨侧面、腹面、背面及下颌侧面的形态进行了初步研究.结果表明,主成分分析和判别函数分析显示头骨侧面和下颌侧面更适宜于区分滇中高原和横断山地区的中缅树鼩;经过薄片样条法分析显示形变多集中在鼻骨和臼齿,这可能与中缅树鼩生存的气候和地理环境相适应;经多维尺度分析显示滇中高原和横断山地区的中缅树鼩头骨有明显差异,这可能与中缅树鼩生活环境的经度和纬度有关.因此,滇中高原和横断山地区的中缅树鼩从头骨形态上可以区分开来,并且差异仅仅发生在种群水平,这可能反映了其对特定生态环境的形态适应.     

限食条件下下丘脑神经肽基因表达量对中缅树鼩体质量调节的影响

为阐明中缅树鼩Tupaia belangeri下丘脑神经肽基因表达量在限食条件下的体质量调节作用,本研究测定了限食(正常摄食量的80%)条件下中缅树鼩的体质量、体脂含量、血清瘦素浓度以及神经肽Y(NPY)、刺鼠相关蛋白(AgRP)、阿片促黑色素原(POMC)和可卡因-安他非明转录调节肽(CART)表达量。结果表明:限食极显著降低中缅树鼩的体质量和体脂含量(P0.01)。对照组和限食组的血清瘦素浓度差异有统计学意义(P0.05),血清瘦素浓度与体脂含量呈正相关。下丘脑神经肽基因NPY、AgRP、POMC和CART表达量差异有高度统计学意义(P0.01)。以上结果表明限食可以降低中缅树鼩的体质量、体脂含量和血清瘦素浓度,调整下丘脑神经肽基因表达量。瘦素通过调节NPY/AgRP和POMC/CART表达量来调节中缅树鼩的体质量。     

DNA条形码技术在中缅树鼩隆安种群和昆明种群鉴定中的研究

目的:本研究利用线粒体细胞色素C氧化酶亚基I基因(CO1)的一段保守区域作为DNA条形码技术的研究序列,探讨DNA条形码技术对中缅树鼩隆安种群和昆明种群进行分类鉴定的可行性。方法:对22只广西隆安树鼩和21只昆明树鼩样本的CO1基因进行PCR扩增、测序,应用MEGA V5软件对序列进行比对及分析其遗传距离,采用NJ法构建系统发育树。结果:中缅树鼩种群中,隆安种群、昆明种群和海南亚种的种内遗传距离为0.00%-0.79%,种群间遗传距离为9.71%-13.59%,中缅树鼩与普通树鼩的种间遗传距离为20.43%-24.11%,存在条形码间隔。系统发育树显示:隆安种群、昆明种群及海南亚种分别聚为一小支,分支置信度高达100%。结论:本研究结果表明DNA条形码技术有助于树鼩种群和亚种的分类鉴定,经CO1基因的测序分析证实广西隆安树鼩和昆明树鼩分属不同的种群。     

中缅树鼩肝在冷适应条件下的产热特征

中缅树鼩(Tupaia belangeri)为东洋界特有小型哺乳动物。本研究测定了中缅树鼩冷驯化实验组(7d、14d、21d、28d)与对照组(0d)中肝线粒体蛋白含量、呼吸状态Ⅲ和状态Ⅳ的变化,探讨中缅树鼩对不同冷环境的适应情况以及肝产热的机理。结果表明,在冷驯化条件下,中缅树鼩产热显著增加,与对照组相比,实验组肝总蛋白含量、肝线粒体蛋白含量、呼吸状态Ⅲ和状态Ⅳ有着显著的提高,在28d后分别增加了39.9%、39.3%、84.9%、181.1%,因此,肝在中缅树鼩冷适应产热过程中具有重要的作用。同时,从生理生态方面为树鼩的岛屿起源学说提供依据。     

基于细胞色素氧化酶亚基Ⅰ基因的中缅树鼩地理种群遗传分化

中缅树鼩广泛分布于东南亚地区,在我国主要分布于西南地区及海南岛。本研究以中国10个地理种群共112只中缅树鼩为研究对象, PCR扩增得到1398 bp的细胞色素氧化酶亚基Ⅰ（COⅠ）基因序列,并对其进行分析。 结果显示：112个中缅树鼩COⅠ基因共定义了64个单倍型,其单倍型多样性（Hd）平均值为0.9730,核苷酸多样性（Pi ）平均值为0.04494;AMOVA方差分析显示种群间的变异占总变异的93.09%,说明中缅树鼩地理种群变异主要发生在种群间;整体遗传分化固定指数（F_ST）为0.93091,说明各地理种群中缅树鼩已出现明显的遗传分化;结合中性检验与碱基错配分布图结果表明中缅树鼩在历史进程中未经历过种群扩张现象; 基于单倍型构建的系统进化树与NETWORK网络图显示10个地理种群的中缅树鼩聚为4支：海南种群一支,大新种群一支,片马种群一支,其他种群一支。结果表明：中国不同地理种群的中缅树鼩具有较高的遗传多样性,各地理种群间已经出现了较为明显的遗传分化,地理阻隔作用可能是其分化的主要原因。     

基于细胞色素b基因探讨昆明禄劝地区树鼩的分类意义

通过对云南省昆明市禄劝地区30只中缅树鼩(Tupaia belangeri)细胞色素b(Cytochrome b,Cyt b)基因全序列(1 140 bp)的遗传分析,对禄劝地区中缅树鼩;Cyt b基因特征进行初步探讨.结果表明,禄劝地区中缅树鼩Cyt b基因包含32个核苷酸变异位点,占全序列的2.81%,其转换/颠换为10.4.基于Cyt b基因的遗传距离构建的分子系统树显示,本实验研究的中缅树鼩是一个独立的分类单位而非普通树鼩(T.glis)的亚种;同时显示攀鼩目(Scandentia)与皮翼目(Dermoptera)亲缘关系较近.     

中缅树鼩头骨及下臼齿几何形态与环境的关系

共选取分布于云南、贵州和广西地区的190个中缅树鼩头骨和32颗臼齿(第二、第三下臼齿),运用几何形态测量法对其头骨侧面、腹面、背面及下颌侧面的形态特征进行主成分分析、薄片样条分析,同时运用统计软件SPSS15.0对头骨和臼齿进行多维尺度分析,以探讨头骨及臼齿形态与环境之间的关系.结果表明,头骨侧面在研究多种群头骨形态时更适宜,更有参考价值;经过薄片样条法分析显示形变多集中在鼻骨和臼齿,这可能与中缅树鼩生存的气候和地理环境相适应;经多维尺度分析显示云南、贵州和广西地区的中缅树鼩头骨具有明显的地区差异,同时,对臼齿的分析显示,臼齿形态具有地区差异,这种变化可能与中缅树鼩生活环境的经度和纬度有关.综上,云南、贵州和广西地区的中缅树鼩的头骨具有形态变异,并且差异仅仅发生在种群水平,这可能反映了其对特定生态环境的形态适应.     

高脂高糖食物诱导中缅树鼩肥胖的研究

为了研究高糖高脂食物诱导中缅树鼩肥胖的模型,在高脂高糖食物诱导条件下进行了中缅树鼩的血液指标测定、脂肪组织流式细胞分析和正电子发射计算机断层显像(positron emission tomography/computed tomography,PET/CT)扫描成像研究。结果表明:在28 d时中缅树鼩白色脂肪组织(white adipose tissue,WAT)质量是对照组的3.7倍,49 d时WAT质量是对照组的6.4倍;49 d时实验组树鼩的甘油三酯(triglyceride,TG)和高密度脂蛋白(high density lipoprotein,HDL)含量与对照组树鼩差异显著,血糖(blood glucose)水平、总胆固醇(total cholesterol,CHOL)和血小板(blood platelet,PLT)含量与对照组树鼩差异极显著;此外,食物诱导28 d组中缅树鼩腹部WAT和褐色脂肪组织(brown adipose tissue,BAT)的18氟-氟化脱氧葡萄糖(18-fluoro-fluorodeoxyglucose,~(18)F-FDG)吸收60 min后,平均辐射值(standardized uptake value,SUV)显著高于对照组,49 d时有所下降。流式细胞分析结果表明,28 d时WAT和BAT中R2群标记阳性显著增加,而49 d后有所降低。以上结果表明:食物诱导可以使中缅树鼩产生肥胖并显著增加中缅树鼩WAT的质量,中缅树鼩在28 d时会出现肥胖抵抗的现象,但是随着时间的延长,该抵抗现象逐渐减少。     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000     

Coiled-coil nanomechanics and uncoiling and unfolding of the superhelix and alpha-helices of myosin

The nanomechanical properties of the coiled-coils of myosin are fundamentally important in understanding muscle assembly and contraction. Force spectra of single molecules of double-headed myosin, single-headed myosin, and coiled-coil tail fragments were acquired with an atomic force microscope and displayed characteristic triphasic force-distance responses to stretch: a rise phase (R) and a plateau phase (P) and an exponential phase (E). The R and P phases arise mainly from the stretching of the coiled-coils, with the hinge region being the main contributor to the rise phase at low force. Only the E phase was analyzable by the worm-like chain model of polymer elasticity. Restrained molecular mechanics simulations on an existing x-ray structure of scallop S2 yielded force spectra with either two or three phases, depending on the mode of stretch. It revealed that coiled-coil chains separate completely near the end of the P phase and the stretching of the unfolded chains gives rise to the E phase. Extensive conformational searching yielded a P phase force near 40 pN that agreed well with the experimental value. We suggest that the flexible and elastic S2 region, particularly the hinge region, may undergo force-induced unfolding and extend reversibly during actomyosin powerstroke.     

Synthesis of hapten and conjugates of coumestrol and development of immunoassay

3-O-Carboxymethylcoumestrol was prepared as the hapten for immunoassay by a partial alkylation of coumestrol with ethyl chloroacetate in acetone alkalized with potassium carbonate. 3-O-Ethoxycarbonylmethylcoumestrol was separated by column chromatography and finally was hydrolyzed with formic acid. 1H and 13C NMR data (APT, COSY, HMQC, and HMBC) revealed that the reaction was regioselective, as 3-O-ethoxycarboxymethylcoumestrol was the only monosubstituted derivative. The hapten was then conjugated to bovine serum albumin and used for immunization of rabbits. A radioimmunoassay (RIA) system was established based on the polyclonal antiserum and a 125I-labeled hapten-tyrosine methyl ester conjugate as the radioligand. Parameters of the RIA: sensitivity: 12 pg per tube, 50% intercept: 140 pg per tube, working range: 20-4000 pg per tube. The cross-reactivity of a panel isoflavonoid and lignan phytoestrogens was either negligible (e.g. formononetin 0.07%; biochanin A 0.06%) or not detectable at all. The major immunoreactive peak in HPLC fractions from an alfalfa extract had the same retention time as coumestrol standard and represented 94.8% of the signal. The remaining 5.2% of immunoreactivity was distributed between five minor peaks. We conclude that after the validation for particular matrices, the method will be a useful tool for analysis of coumestrol, especially in low volume and low concentration samples.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。     

Effect of soil salinity and water stresses on growth and content of nitrogen,chloride and phosphate of wheat and triticale

Summary Three wheats and one triticale were grown, up to flowering stage, in pots on calcareous soil adjusted to a range of salinities (S₁=3.5, S₂=6, S₃=8.5, and S₄=11 mmhos/cm, 20°C, soilpaste extract) by adding solution consisting of 3∶2∶1 of Na-, Ca- and Mg chlorides in chemical equivalent amounts. Moisture in the pots was kept at 100% (W₁), 40% (W₂) and 20% (W₃) of the available water. The vegetative growth, nitrogen and phosphate were affected by S and W treatments, chloride was affected only by S. The interaction S×W affected only dry weight. Varietal effect was observed between wheat as a group and triticale. Multiple quadratic regression equations of these properties on salinity and water revealed that the higher the available water the wider the range of tolerable salinity. Triticale was relatively more tolerant to water stress. Salinity increases Cl and decreases N, whereas water stress enhances N accumulation to a certain extent. However, in triticale at S₃ and S₄ the effect of water stress on N was overshadowed by the excessive salinity. This did not occur for the wheat (Florence). P trends were described. R² for P was low (0.7435–0.3603) which made interpretations rather difficult.     

放牧对牧草光合作用、呼吸作用和氮、碳吸收与转运的影响

研究放牧对草地植物生理活动的影响,对于揭示草地放牧演替的生理机制有重要意义.大量研究表明,家畜放牧对牧草光合作用、呼吸作用以及C和N吸收与转运的影响,可以分为生理伤害和生理恢复2个阶段.放牧通过改变草地冠层结构影响牧草光合作用,净光合作用速率短期内迅速下降,随着叶面积指数增加又逐渐上升,呼吸作用有相似的变化趋势.牧草放牧后再生长所需的C和N最初主要来自根系和留茬中的贮藏物质,此后随着牧草生长恢复逐渐由同化作用供给,C代谢与土壤N水平负相关.放牧后牧草生理活动变化与牧草遗传特性、种间竞争、家畜放牧特征、非生物环境等因素密切相关.