十三碳二元羧酸发酵技术的研究

以一株热带假丝酵母菌(Candida tropicalis) SP1为出发菌株,经紫外线反复诱变获取一株难以同化烷烃的突变株SPUV56,摇瓶培养5d平均产酸量达72g/L,较出发菌株提高了1.25倍,并利用突变株SPUV56在137L自控罐上扩试,补加醋酸盐发酵,144h产酸量达153g/L,比不加醋酸盐发酵提高了29.7%。采用提 高搅拌混合效果和低溶解氧发酵过程控制方法,可有效地提高菌体的产酸能力,在20m3发酵罐中发酵生产十三碳二元羧酸,总培养时间144h,产酸量可达172g/L,放罐体积15.0m3,产量为2.25t。     

唐家河自然保护区大熊猫种群数量及栖息地的移动

本文根据大熊猫季节性活动规律特点,应用海拔高度路线调查法,全面收集唐家河自然保护区大熊猫的比较新鲜粪便,计获１２４个粪样材料,提取ＤＮＡ作ＤＮＡ指纹图,经微机检测,保护区有大熊猫３７只,个体数量与粪样采集量之比为１∶３３５１４,有６个家系,雌兽２１只,雄兽１６只。并搞清了保护区生态环境质量,箭竹遭受病虫害的程度,洪水对栖息地的影响,特别对引起大熊猫数量减少及栖息地移动等原因,作了较为详细的分析。     

Comprehensive evaluation of multiple cropping systems on upland red soil

According to the principles and methods of ecology and system engineering, we set up an evaluation indicator system for multi-component and multiple cropping systems, evaluated the comprehensive benefits of multi-component and multiple cropping systems using grey relation clustering analysis and screened out the optimized model based on research done in the upland red soil in Jiangxi Agricultural University from 1984 to 2004. The results show that the grey relation degree of “cabbage/potato/maize — sesame” was the highest among 23 multi-component and multiple cropping systems and was clustered into the optimized system. This indicates that “cabbage/potato/maize — sesame” can bring the best social, economic and ecological benefits, increase product yield and farmers’ income and promote sustainable development of agricultural production. Therefore, it is suitable for promotion on upland red soil. The grey relation degree of “canola/Chinese milk vetch/maize/mung bean/maize” was second, which is suitable for implementation at the city outskirts. In conclusion, these two planting patterns are expected to play important roles in the reconstruction of the planting structure and optimization of the planting patterns on upland red soil. __________ Translated from Acta Ecologica Sinica, 2006, 26(8): 2532–2539 [译自: 生态学报]     

青岛近海及其临近海域冬季微微型浮游植物的分布

微微型浮游植物（0.2～2.0 μm） 是海水中最小的自养浮游生物, 在世界各海域广泛分布, 并在海洋有机物质循环中起着非常重要的作用.利用荧光显微技术对青岛近海及其邻近海域冬季微微型浮游植物丰度进行了调查,研究了微微型浮游植物的空间变化和昼夜变化的特征, 并分析了微微型浮游植物丰度与环境因子的相关性. 结果显示, 冬季该海域以富含藻红素（Phycoerythrin-rich, PE）的聚球藻(Synechococcus, Syn)细胞占优势,微微型真核藻类（Picoeukaryote, Euk）次之,而富含藻蓝素（Phycocyanin-rich, PC）的聚球藻细胞数量很低, 未发现原绿球藻（Prochlorococcus, Pro）的存在.Syn 的变化范围为8.97×10³～1.95×10⁵ cells·ml^-1, 平均4.67×10⁴ cells·ml^-1; Euk的变化范围为1.95×10²～1.01×10⁴ cells·ml^-1, 平均2.39×10³ cells·ml^-1. Syn丰度在胶南以南海域出现高值区域, 在即墨海域和崂山东南海域出现低值区域. Euk丰度在日照海域出现高值区域; 崂山海域为低值区域; 各水层Syn和Euk丰度均无明显差异(P﹥0.05). 对胶州湾中部连续站4层水体的24 h昼夜连续变化进行观测发现, Syn、Euk丰度都有明显昼夜波动.相关性分析表明： Syn与温度、 电导率呈正相关, 与溶氧浓度呈显著负相关; Euk与盐度和溶氧浓度呈显著负相关. 微微型浮游植物对总浮游植物生物量的贡献约为20％.     

Inhibition of mitochondrial complex I improves glucose metabolism independently of AMPK activation

Accumulating evidences showed metformin and berberine, well‐known glucose‐lowering agents, were able to inhibit mitochondrial electron transport chain at complex I. In this study, we aimed to explore the antihyperglycaemic effect of complex I inhibition. Rotenone, amobarbital and gene silence of NDUFA13 were used to inhibit complex I. Intraperitoneal glucose tolerance test and insulin tolerance test were performed in db/db mice. Lactate release and glucose consumption were measured to investigate glucose metabolism in HepG2 hepatocytes and C2C12 myotubes. Glucose output was measured in primary hepatocytes. Compound C and adenoviruses expressing dominant negative AMP‐activated protein kinase (AMPK) α1/2 were exploited to inactivate AMPK pathway. Cellular NAD⁺/NADH ratio was assayed to evaluate energy transforming and redox state. Rotenone ameliorated hyperglycaemia and insulin resistance in db/db mice. It induced glucose consumption and glycolysis and reduced hepatic glucose output. Rotenone also activated AMPK. Furthermore, it remained effective with AMPK inactivation. The enhanced glycolysis and repressed gluconeogenesis correlated with a reduction in cellular NAD⁺/NADH ratio, which resulted from complex I suppression. Amobarbital, another representative complex I inhibitor, stimulated glucose consumption and decreased hepatic glucose output in vitro, too. Similar changes were observed while expression of NDUFA13, a subunit of complex I, was knocked down with gene silencing. These findings reveal mitochondrial complex I emerges as a key drug target for diabetes treatment. Inhibition of complex I improves glucose homoeostasis via non‐AMPK pathway, which may relate to the suppression of the cellular NAD⁺/NADH ratio.     

HOPS复合体蛋白在细胞自噬过程中的功能研究

自噬(autophagy)是一种溶酶体依赖性的细胞内降解途径,其主要功能是将生物大分子(蛋白质、多糖等)或细胞器(线粒体等)回收至溶酶体中并将其降解为单糖、氨基酸等小分子以重复利用。发现HOPS复合体中的两个基因vps39和vps41的缺失会导致酵母内GFP-ATG8大量积累。进一步研究表明,积累的原因是GFP-ATG8与液泡不能发生融合。而在HOPS复合体中的另外两个基因vps16和vps18缺失的情况下,自噬融合没有受到影响;在vps16和vps18双敲除的菌株中,自噬融合同样没有受到影响。该实验结果为理解HOPS复合体的功能和自噬体与液泡融合的过程提供了新的线索。     

应用微卫星标记对雌核发育银鲫的遗传多样性初探

利用Crooijmans et al.(1997)分离的包含CA重复单元的普通鲤鱼（Cyprinus carpino.L)的8个微卫星DNA标记,对银鲫（Carassius auratus gibelio Bloch)的5个不同雌核发育系的24尾个体进行PCR扩增。分析电泳结果发现,除MFW28未能在银鲫中稳定地扩增出相应的同源序列,其余的7对引物扩增的重复性和稳定性都很好,随引物不同,各等位基因数为1-14个,大小在100-506bp。在MFW1、MFW4、MFW19、MFW20、MFW23和MFW246个微卫星的扩增图谱中,不同的雌核发育系扩增出各自独特的图谱,而同一系内的不同个体间具有高度的遗传同质性,但仍然在个别个体中检测到少量的多态片段。不同系间的扩增图谱呈现出高度的遗传异质性,共鉴定出23个可以用于有效区分5个不同雌核发育系的分子标记。这5个微卫星标记反映了银鲫5个雌核发育系间的相互亲缘关系,其中P和A系同属一个雌核发育系,F系起源于E系,A、D和E系可能分别独立地起源于不同的杂交事件,鉴定的微卫星分子标记为进行银鲫群体遗传学和进化遗传学研究,以及银鲫的分子标记育种和进行基因组作图提供了理想的工具。     

低频脉冲电场对PC12细胞酪氨酸羟化酶和多巴胺合成的影响

运用Western印迹和HPLC分别测定不同时间电场刺激和刺激后不同培养时间条件下,PC12细胞内酪氨酸羟化酶(TH)和细胞培养液中多巴胺(DA)含量的变化。结果显示,受到短时间(5、10min)脉冲电场刺激的PC12细胞,经较短时间(2天)的培养后,细胞内TH的含量和培养液中DA的含量均比对照组有所提高,但随着培养时间的延长(3~5天),TH和DA的含量均明显下降。然而,长时间(15、20、30min)脉冲电场刺激组则先表现为TH和DA的合成受到抑制,但随着培养时间的延长,其合成则被逐渐激活。采用蛋白激酶A(PKA)特异性抑制剂H-89和有丝分裂原活化蛋白激酶的激酶(MEK1/2)特异性抑制剂U0126,研究脉冲电场刺激所激活的与TH和DA合成相关的信号通路。结果表明,在没有神经生长因子(NGF)存在的情况下,PC12细胞主要通过PKA通路来激活TH的合成,低频脉冲电场刺激也主要激活PKA通路,因为抑制这条信号通路能显著抑制电场刺激所诱导的TH合成。     

真盐生植物盐地碱蓬根系边缘细胞在耐盐中的作用初探

以黄河三角洲潮间带盐地碱蓬种子生成的幼苗为材料,研究了NaCl胁迫对盐地碱蓬生长与根系边缘细胞的影响。盐地碱蓬的第一个边缘细胞几乎与根尖同步产生,当根长达到13mm时,边缘细胞数目达到最大值。NaCl胁迫抑制边缘细胞的活性,但低浓度的NaCl处理增加边缘细胞的数目。低浓度NaCl处理时果胶甲基酯酶（PME）的活性比对照有明显增加,超氧化物歧化酶（SOD）活性随着NaCl浓度的增加呈现先上升后下降的趋势,低浓度NaCl可以增加盐地碱蓬根内过氧化氢酶（CAT）的活性,NaCl处理时间和处理浓度都对过氧化物酶（POD）活性的影响不明显。这些结果表明,盐地碱蓬至少部分通过增加调控活性氧（ROS）水平增加PME活性及根系边缘细胞数目来抵抗NaCl胁迫。