玉米质体型AGPase小亚基和糖酵解关键酶PPDK1互作关系的研究

【目的】利用双分子荧光互补(BiFC)技术,在烟草叶肉细胞中分析玉米ADP-葡萄糖焦磷酸化酶质体型小亚基(AGPase-bt2)与丙酮酸磷酸双激酶(PPDK1)之间的相互作用。【方法】构建326-CYCHA-AGPase-bt2和326-CYNEE-PPDK1双分子荧光表达载体,转化农杆菌EHA105,瞬时浸染烟草叶肉细胞,激光共聚焦显微镜下观察AGPase-bt2和PPDK1的相互作用。【结果】双酶切试验表明,326-CYCHA-AGPase-bt2、326-CYNEE-PPDK1载体构建正确;PCR结果证实,植物表达载体成功转化到农杆菌EHA105中;浸染烟草叶片后,AGPase-bt2和PPDK1在叶肉细胞中高效表达,出现BiFC荧光信号。【结论】AGPase-bt2和PPDK1在植物细胞内存在真实互作关系。     

灌浆结实期弱光对水稻籽粒淀粉积累及相关酶活性的影响

 选用IR72（籼稻）和日本晴（粳稻）为材料，在开花后进行遮光处理，对弱光条件下籽粒淀粉和直链淀粉含量的动态变化及相关酶的活性进行了研究。在弱光条件下，两品种籽粒的淀粉含量减少，直链淀粉含量下降，蔗糖含量降低，ADPG焦磷酸化酶活性的变化较小，可溶性淀粉合成酶和颗粒结合型淀粉合成酶活性减弱，可溶性淀粉分支酶Q酶和颗粒结合型淀粉分支酶活性增强，淀粉去分支酶活性因品种而异，IR72表现为减弱，日本晴则为增强。相关分析表明，遮光下蔗糖输入量的减少量和淀粉合成量的下降量呈显著正相关；ADPG焦磷酸化酶和淀粉分支酶活性与淀粉积累速率呈显著正相关。分析指出，遮光下淀粉合成酶活性的降低与淀粉合成量的下降有关，淀粉分支酶活性的升高是直链淀粉占淀粉总量的比率减少的重要原因。     

Cassava starch biosynthesis: new avenues for modifying starch quantity and quality

The genes coding for the main enzymes involved in cassava starch biosynthesis have been cloned and characterised. Molecular analysis of these genes revealed high amino acid sequence homology with other cloned genes from starch forming plant species. Use of these genes to modify the pathway of starch biosynthesis in cassava has become possible with the advent of a reproducible transformation and regeneration protocol for cassava. This would enable the development of new cassava cultivars able to produce starches with different physico-chemical properties and uses. This revised version was published online in July 2006 with corrections to the Cover Date.     

Impact of cycloheximide addition on adenylates in soil

Cycloheximide inhibits specifically the ribosomal protein synthesis of eukaryotic cells, i.e. the metabolism of soil fungi. We measured cycloheximide effects on adenylates in 20 different soils (0-10 cm depth) from arable, grass and forest land with a large variety of soil properties. The aims were (1) to assess the interactions between cycloheximide effects and soil properties and (2) to prove the relationship between cycloheximide effects on ATP and the ergosterol-to-microbial biomass C ratio, which is an indicator for the fungal proportion of the total microbial biomass. The adenylates ATP, ADP and AMP were measured 6 h after adding either 10 mg cycloheximide per gram soil in combination with 24 mg talcum per gram soil or 24 mg talcum per gram soil solely. The medians of the relative increases in AMP and ADP were 45 and 25% and the medians of the relative decreases in ATP and adenylates were −36 and −12%. These changes in adenylate composition lead to a cycloheximide-induced relative decrease in the adenylate energy charge level of 15%. The relative decrease in ATP content after cycloheximide addition was significantly correlated with the ATP-to-microbial biomass C ratio, but not with the ergosterol-to-microbial biomass C ratio. The absolute increase in ADP and the absolute decrease in ATP were affected by the clay content according to principal component analysis. The reduction of the ATP-to-microbial biomass C ratio indicates that this ratio had the potential of being an important ecotoxicological indicator of direct toxic effects of organic pollutants on soil microorganisms.     

节杆菌AD26的分离鉴定及其与假单胞菌ADP对阿特拉津的联合降解

用富集培养法,从农药厂的工业废水中分离到高效降解除草剂阿特拉津的AD26菌株,通过16SrRNA基因序列分析,该菌株被鉴定为节杆菌（Arthrobacter sp.）。降解基因的PCR分析表明,AD26含有阿特拉津降解基因trzN和atzBC,它能以阿特拉津为唯一氮源、蔗糖或柠檬酸钠为碳源生长,将阿特拉津降解成氰尿酸,降解速度快但降解不完全。假单胞菌（Pseudomonas sp.）ADP是Wackett实验室分离的阿特拉津降解菌株,含有阿特拉津降解基因atzABCDEF,能以阿特拉津为唯一氮源、柠檬酸钠为碳源（不能以蔗糖为碳源）生长,将阿特拉津降解成NH3和CO2,降解完全但降解速度慢。在阿特拉津浓度为200mg·L^-1的无机盐培养基中进行的AD26和ADP混合培养表明,它们对阿特拉津的降解发生了互补和增强作用,两个菌株能在以阿特拉津为唯一氮源、蔗糖为碳源的培养基中生长,而且生长和降解速率都好于单个菌株,培养72h后阿特拉津去除率达到99.9%,其中76.7%的阿特拉津被降解成NH3和CO2。这表明由节杆菌AD26和假单胞菌ADP组成的混合菌株在阿特拉津废水处理和污染土壤的生物修复中有很好的应用潜力。     

能量物质在体外对大鼠快肌和慢肌卫星细胞增殖与抗氧化机能研究

探讨体外能量物质对SD大鼠快肌(胫前肌)与慢肌(股中间肌)卫星细胞增殖功能和H2O2化学损伤修复研究。采用快、慢肌卫星细胞传代培养及分离纯化,四唑盐(MTT)比色法、DAPI荧光染色与免疫荧光抗体检测。结果表明:1与正常组比较,低剂量或中等剂量能量物质对快肌与慢肌卫星细胞有增殖机能,各剂量作用时间24h48h72h,其中以ATP溶液1.25mg/mL,ADP溶液1.25mg/mL,AMP溶液2.5mg/mL剂量促进快、慢肌卫星细胞增殖效果最显著;2 MTT比色法检测ATP、ADP及AMP修复损伤快、慢肌卫星细胞明显低于正常组卫星细胞而高于纯H2O2损伤卫星细胞。且修复效果表现为:ATP溶液以1.25mg/mL,ADP溶液为2.5mg/mL,AMP溶液为2.5mg/mL修复最佳;3模型组Bax抗体细胞数102.5±5.7,Bcl-2抗体细胞29.6±3.7;ATP、ADP及AMP溶液保护组Bax抗体细胞数67.3±4.9~72.8±5.4范围,而Bcl-2抗体细胞在69.8±5.1~78.5±4.9范围之内。慢肌股中间肌卫星细胞各组检测结果与其类似。     

高效液相色谱法测定龙眼果实中ATP、ADP、AMP的含量

摸索了一种基于高效液相色谱仪快速测定龙眼果实腺苷三磷酸(ATP)、腺苷二磷酸(ADP)、腺苷一磷酸(AMP)含量的方法.该方法利用磷酸盐缓冲液(pH7.0)作为单一流动相,流速设定为1 mL/min,停止时间13 min,检测波长254 nm,无参比波长.ATP、ADP和AMP浓度在5～50 mg/L范围内与相应的色谱峰面积呈完美的直线关系(决定系数R2均为1.000),回收率95.2％ ～97.5％,保留时间分别为8.651、9.844和11.457 min.该方法测定结果表明,果实腺苷含量水平分别为30.0、6.5和2.0μg/g,能荷值为0.86.     

Farmers’ Assessment of Extension Services in Ogun State,Nigeria

This study examined farmers’ assessment of extension services in Ogun State, Nigeria. One hundred and twenty respondents were randomly selected and an interview schedule was used to collect data. Frequency distribution, percentages, chi-square, and Pearson Product Moment Correlation were used to analyze the data. The results revealed that more than half of the farmers (55%) fall within the age range of 40–59, most are female (66.7%), 70% have some level of formal education, and the majority (80%) had more than 10 years of experience with more than one farm location. A high percentage (90.0%) had frequent contact with extension agents, but less than half (49.2%) acknowledged extension services to be effective. Extension agents should be equipped with necessary materials and training in order to improve their services.     

高效液相色谱法测定大鼠比目鱼肌和内侧腓肠肌内ATP·ADP及AMP含量的研究

[目的]比较成年大鼠比目鱼肌和内侧腓肠肌内ATP、ADP及AMP的含量。[方法]采用大体解剖法取出大鼠的比目鱼肌和内侧腓肠肌,置于液氮内研磨成粉末,再分别加入EDTA和HClO4、NaHCO3,经高速冷冻离心取出上清液,用高效液相色谱仪测定。[结果]慢肌(比目鱼肌)ADP的含量多于快肌(内侧腓肠肌),慢肌(比目鱼肌)ATP、AMP的含量少于快肌(内侧腓肠肌)。[结论]使用高效液相色谱仪可以检测出快慢肌的差异。