AtDME1化学诱导表达载体的构建及其瞬时表达

以拟南芥去甲基化转移酶基因DME1为目的基因,采用酶切、连接和重组反应的方法构建了以17-β-雌二醇为诱导剂的植物表达载体p ER8-DME1。将其转入农杆菌LBA4404菌株中,通过烟草瞬时表达技术,对该载体的诱导表达特性进行了研究。q PCR结果表明,较低浓度的17-β-雌二醇处理能够有效调控p ER8-DME1中目的基因的表达,诱导处理后目的基因表达量逐渐升高,在12 h后达到最高,之后缓慢下降。该载体的成功构建为深入研究DNA去甲基化与植物的生殖发育调控奠定了基础。     

红色和橙黄色果肉西瓜番茄红素和β–胡萝卜素代谢基因的表达分析

以橙黄色果肉西瓜‘WM-Clr-2’及红色果肉西瓜‘PI 179881’为试材,采用HPLC法分析其番茄红素及β–胡萝卜素在7个发育时期的积累情况,同时采用q RT-PCR技术分析番茄红素和β–胡萝卜素代谢途径中8个关键酶基因(PSY1、PSY2、PDS、ZDS、CRTISO、LCYB、NCED1和NCED7)的表达量。结果显示,随着果实的发育,番茄红素和β–胡萝卜素含量均不断增加,且在同一时期,红色果肉‘PI 179881’中的含量始终大于‘WM-Clr-2’。q RT-PCR结果显示PSY1在‘PI 179881’中的表达量高于‘WM-Clr-2’,LCYB和NCED1在‘WM-Clr-2’中的表达量高于红色果肉‘PI 179881’。分析番茄红素及β–胡萝卜素的积累规律与相关基因表达规律说明PSY1的大幅上调表达,LCYB的下调表达及NCED1的低表达可能是红色果肉‘PI 179881’中番茄红素和β–胡萝卜素大量积累的主要原因。PSY1的低表达水平,LCYB和NCED1的高表达水平可能是橙黄色果肉‘WM-Clr-2’中番茄红素及β–胡萝卜素积累量少的主要原因。     

Aβ1-42淀粉样蛋白在大肠杆菌中的可溶性表达

本研究中用融合标签技术通过单质粒转化和双质粒转化分别促进Aβ1-42淀粉样蛋白的可溶性表达。构建重组载体pET（yd-b42）、pET（Msb-b42）、pET（Od-b42）、pAY-sls[ydb42]、pAY-sls[tydb42],并在大肠杆菌中表达,通过SDS-PAGE验证分析。标签yd、Msb、Od、tyd都能很好的促进Aβ1-42淀粉样蛋白可溶性表达,其中yd、tyd的促溶效果最好。Aβ1-42淀粉样蛋白能在大肠杆菌中可溶性表达。为阿尔茨海默病的治疗提供进一步理论基础。     

磷脂酶Dβ在拟南芥低温信号中的转导作用

磷脂酶D（PLD）不仅是植物中一类主要的磷脂水解酶,而且是一类重要的跨膜信号转导酶类。PLD的磷脂降解功能和信号转导功能均影响植物的抗冻性。本研究以PLDβ基因被敲除的拟南芥突变体及其野生型植株为材料,进行低温驯化和冻害胁迫处理,并分析其作用途径。结果表明,PLDβ基因介导低温信号转导作用,参与渗透调节途径中脯氨酸的调控和抗氧化系统中过氧化氢酶（CAT）活性的调控,并且与低温信号激素ABA不在同一条信号转导途径。本研究为探索通过调控PLD的活性提高植物抗冻性提供了新的途径,并为深入揭示植物的抗冻机理以及磷脂信号转导机制提供实验支持。     

繁殖基因(ESR、LH、LHR、PRLR)编码区生物信息学分析

为了研究各物种ESR、LH、LHR、PRLR基因进化的特点,从GenBank下载了人、毛猩猩、黑猩猩、猕猴、欧洲兔、野猪、黄牛、大熊猫、小家鼠、仓鼠、褐家鼠11个物种的139条编码区序列(CDS),采用生物信息学方法对4种基因进行分析。结果表明,4种基因均存在较丰富的遗传多样性。其中ESR基因单位长度多态位点数为0.330 7,单位长度突变位点数为0.399 8,核苷酸多样性最低,同义替换率相对最高,平均遗传距离最小。LH基因单位长度多态位点数(0.484 8)与单位长度总突变位点(0.618 3)均最高,核苷酸多样性和非同义替换率相对最高。依据4种基因遗传距离构建的聚类图的拓扑结构存在差异,说明4种繁殖基因进化速率存在差异,ESR基因进化速率相对较慢,LH基因相对较快。     

不结球白菜感染菌核病后4个生理生化指标分析

选用4个对菌核病具有不同抗性的不结球白菜品种003-R-6、Y3-R-4、109-S-9和003-S-7为试验材料,于四叶一心期接种核盘菌,采用苗期琼脂块叶片接种法测定各品种对菌核病的抗性,并分别测定接种前后叶片中4个生理生化指标。结果表明,苗期琼脂块叶片接种法与田间自然诱发鉴定结果相一致,可有效反映抗、感品种的差异。在核盘菌影响下,4个抗、感病品种中不结球白菜的丙二醛(MDA)含量、游离氨基酸(FAA)总量、几丁质酶活性和β-1,3-葡聚糖酶活性均有所变化。其中丙二醛(MDA)含量、游离氨基酸(FAA)总量表现为感病品种增加幅度大于抗病品种,而几丁质酶活性、β-1,3-葡聚糖酶活性均表现为抗病品种增加幅度大于感病品种,说明各指标与品种抗病性强弱存在一定的相关性。     

Selected Soil Enzyme Activities,Soil Microbial Biomass Carbon,and Root Yield as Influenced by Organic Production Systems in Sweet Potato

A field experiment was conducted on Typic Rhedustalfs to determine the effects of various organic production systems. Results revealed that the soil microbial biomass carbon (SMBC) content was greatest with the application of 100% N through farmyard manure. The ratio index value of biofertilizer along with 50% N through any one of the organic sources were greater than 100% N through green leaf manure?/?vermicompost, integrated use of manure and fertilizer (conventional production system), and control (traditional system of production). Soil enzymes varied with the production systems. The urease, phosphatase, and β-glucosidase activities were more with greater nitrogen, phosphorus, and organic-matter treatments, respectively. The SMBC, soil enzymes, and microbial activity were very responsive to organic production systems, but their levels and activities were not reflected in sweet potato root yield.     

高活性β-葡萄糖苷酶丹贝发酵工艺研究

本文以大豆为原料,少孢根霉为发酵菌株,在研究蒸煮时间、接种量和发酵时间3个单因素条件对丹贝中β-葡萄糖苷酶活性影响的基础上,应用Box-behnken中心组合实验方法,以β-葡萄糖苷酶活性为响应值,进一步优化丹贝的生产工艺,结果表明:蒸煮时间为20min,接种量为1.50%,发酵时间为36h的条件下β-葡萄糖苷酶活性最高,为345.63U,这可为丹贝的工业化生产提供一定的理论依据。     

热烫方式对胡萝卜浆品质的影响及工艺优化

为提高胡萝卜制浆的品质,以新鲜东方六寸参胡萝卜为原料,采用高效液相色谱法测定胡萝卜浆中β-胡萝卜素含量。比较了热水热烫、蒸汽热烫、维生素C热烫、柠檬酸热烫等4种不同的热烫方式对胡萝卜浆中β-胡萝卜素含量的影响,并用Box-Behnken响应面设计法优化热烫处理的工艺条件。结果表明,柠檬酸热烫为最佳热烫方式,优化得到的柠檬酸热烫工艺条件为:柠檬酸质量分数1.4%、热烫时间18min、热烫温度84℃。经验证在此条件下,β-胡萝卜素质量分数23.59mg/kg,与模型预测值吻合,建立的模型可用于工艺预测。因此,本研究结果可为改善胡萝卜制浆的品质提供技术依据。     

Molecular aspects of cereal β-glucan functionality: Physical properties,technological applications and physiological effects

Cereals β-glucans are linear homopolysaccharides of consecutively linked (1→4)-β-d-glucosyl residues (i.e. oligomeric cellulose segments) that are separated by single (1→3)-linkages. β-Glucans display all the functional properties of viscous and gel forming food hydrocolloids combined with all the physiological properties of dietary fibres. This review focuses on the relationships between the molecular–structural characteristics of β-glucans and their physicochemical properties in aqueous dispersions and in food systems as well as their physiological functions in the gastro-intestinal tract. The physical properties of β-glucans, such as solubility and rheological behaviour in the solution and gel states, are controlled by their molecular features, such as their distribution of cellulosic oligomers, their linkage pattern and their molecular weight as well as by temperature and concentration. The technological and nutritional functionality of β-glucans is often related to their rheological behaviour. Incorporation of β-glucans into various products (bread, muffins, pasta, noodles, salad dressings, beverages, soups, reduced-fat dairy and meat products) showed that attributes, such as breadmaking performance, water binding and emulsion stabilising capacity, thickening ability, texture, and appearance appear to be related to the concentration, molecular weight and structure of the polysaccharide. The health benefits of β-glucans, such as reducing blood serum cholesterol and regulating blood glucose levels, are also correlated with the amount and molecular weight of the solubilised β-glucans in the gastro-intestinal tract.