超声波对尼罗罗非鱼幼鱼生长的影响

选择尼罗罗非鱼Tilapia nilotica幼鱼(平均体重为1.95 g±0.60 g)为试验对象,用3种不同强度的超声波对其进行辐射刺激,共设3个试验组和1个对照组。经过60 d的超声波辐射养殖,结果表明:各组间体重存在极显著性差异(P<0.01),其中每天受频率为23 kHz、声源声强为200 mW/cm2的连续超声波辐射刺激5 min的试验组鱼,其体重最高,比对照组鱼体重增长约7.58%;但每天受频率为23 kHz、声源声强为600 mW/cm2的连续超声波辐射刺激5 min的鱼体,其体重反而比对照组体重减少约5.80%。各试验组尼罗罗非鱼体重与胃蛋白酶活性之间没有明显关系。当环境声强为1.58~1.60 mW/cm2时,对尼罗罗非鱼幼鱼生长比较有利。     

玉米受伤诱导基因Wip1的启动子克隆及表达分析

【目的】植物蛋白酶抑制剂是抵抗动物摄食和病原侵染的重要防御蛋白。玉米Wip1(wound-induced protein)基因属于Bowman-Birk蛋白酶抑制剂(BBI)家族,了解Wip1(wound-induced protein)的启动子活性和Wip1组织表达特性、受胁迫诱导表达情况和在不同受伤时间下的表达模式,为抗虫基因在植物中应用提供新信息。【方法】以玉米叶片组织的cDNA为模板,采用克隆测序技术获得Wip1启动子序列和该基因的cDNA序列。将所得启动子与植物表达载体p1300连接,构建重组表达载体p1300-Wip1-GUS,并采用冻融法将其转入农杆菌LBA4404中。利用农杆菌介导的瞬时表达体系在玉米愈伤组织中对Wip1启动子的表达活性进行分析。利用热酚法提取玉米相应组织的总RNA,纯化后于含甲醛的变性胶中电泳分离,并将RNA转移到尼龙膜上,用[α-32P]dCTP标记探针的Northern blot技术对Wip1在玉米不同组织、不同胁迫条件、不同受伤时间下的表达模式进行分析。【结果】获得的启动子序列全长1 737 bp,cDNA全长512 bp。将含有重组质粒p1300-Wip1-GUS的农杆菌侵染玉米愈伤组织,3 d后,GUS染色显示侵染部位变蓝,说明所克隆的启动子在玉米愈伤中能启动GUS正常表达。基于Northern blot技术的植物组织表达结果显示,正常情况下,Wip1仅在胚芽鞘中有一定的表达,在受伤后的胚芽鞘、根、茎、叶、雌穗、雄穗、花丝及苞叶中均大量表达;在所设置的胁迫处理时间(2 h)内,Wip1不受缺氧、低温、脱水、PEG、ABA、NaCL和IAA胁迫诱导,仅受伤害胁迫诱导表达;Wip1在玉米叶片受伤胁迫下,玉米叶片受伤2 h时,检测到了Wip1表达,4—24 h时表达量迅速提高且处于持续增加趋势。【结论】玉米Wip1特异地受机械伤害诱导表达;Wip1启动子是一种有效的伤害诱导特异性启动子。     

盐度对仿刺参蛋白摄入量及蛋白酶活性的影响

为研究盐度对仿刺参(Apostichopus japonicus)蛋白摄入量及蛋白酶活性的影响,通过考马斯亮蓝G250染色法间接测定仿刺参蛋白摄入量,福林-酚法测定仿刺参消化道蛋白酶活力。结果表明,在盐度为3.1%时,仿刺参蛋白摄入量最多,消化道蛋白酶活力最强;在盐度2.5%~3.1%范围内,随着盐度的升高,仿刺参蛋白摄入量和消化道蛋白酶活力逐渐升高;在3.1%~3.4%范围内,随着盐度升高,仿刺参蛋白摄入量和消化道酶活力均下降。盐度对仿刺参蛋白摄入量及蛋白酶活性的影响明显。     

枯草芽孢杆菌发酵鱼粉产蛋白酶及其酶解效果

为开发以鱼粉为原料的优质饲料蛋白源以及相关保健食品,开展了用不同鱼粉为氮源发酵枯草芽孢杆菌生产蛋白酶及其酶解效果的研究,比较了不同菌种的发酵效果,确定了最佳发酵时间.通过蛋白酶活性测定和氨基态氮含量测定分析了不同菌种的产酶能力和酶活性大小及其酶解效果.结果表明,用罗非鱼鱼粉和处理过的鳕鱼鱼粉发酵商业化的工业菌种枯草芽孢杆菌产生的酶活性显著高于鲢鱼鱼粉（P〈0.05）,发酵后上清液中的氨基态氮含量也显著高于鲢鱼鱼粉（P〈0.05）,即工业化的枯草芽孢杆菌对不同鱼粉的发酵效果不同,罗非鱼鱼粉和处理过的鳕鱼鱼粉发酵效果显著优于鲢鱼鱼粉.     

月季花瓣丝氨酸蛋白酶基因RhSep1的克隆及生物信息学分析

利用RT-PCR技术,从切花月季品种"Samantha"中克隆丝氨酸蛋白酶基因RhSep1。利用生物信息学技术对所得到的丝氨酸蛋白酶RhSep1序列进行结构与功能预测。结果表明:RhSep1基因全长2 442 bp,开放阅读框编码769个氨基酸,推定该丝氨酸蛋白酶分子质量为80.48 kD。通过NCBI和MEROPS肽酶数据库等对Rh-Sep1进行Protein Blast,发现RhSep1具有肽酶S8₃家族SA的典型结构域Petidase_S8₃（序列为Tyr-108-Leu-341）和PA_subtilisin_like的结构域（序列为Tyr-348-Ile-474）。预测RhSep1可能具有信号肽、明显疏水区和典型跨膜区,同时RhSep1氨基酸序列中存在较多蛋白激酶C和酪蛋白激酶Ⅱ磷酸化位点、豆蔻酰化位点。这些活性位点往往与蛋白的磷酸化、G蛋白相互作用等信号转导事件有关,RhSep1可能存在复杂的蛋白水平调控机制。     

黄绿绿僵菌对马铃薯瓢虫体内脯活性的影响

对感染黄绿绿僵菌后的马铃薯瓢虫幼虫体内几种酶进行测定，结果表明：黄绿绿僵菌对于马铃薯瓢虫幼虫体内的蛋白质含量、蛋白酶、谷胱甘肽-S-转移酶和乙酰胆碱酯酶的活性，都有不同程度的影响，其中对蛋白含量和谷胱甘肽-S-转移酶的影响较大。     

The carboxyterminal processing protease of D1 protein: Herbicidal activity of novel inhibitors of the recombinant and native spinach enzymes

The carboxyterminal processing protease of D1 protein (CtpA) is predicted to be an excellent target for the discovery of a general broad-spectrum herbicide. Directed and random screening of compounds against recombinant spinach CtpA (rCtpA) has led to the discovery of five different chemical classes of inhibitors. Lead compounds from each inhibitor class were investigated for their in vitro effects on the activities of both recombinant and native spinach CtpA. All of the lead compounds have K_i values of less than 50 μM when tested against rCtpA, and all except one showed competitive inhibition. Results from partially purified native CtpA from spinach were similar to those from the recombinant form of the enzyme, thus validating the use of rCtpA in the inhibitor screen. Compounds from three of the classes of CtpA inhibitors show in vivo herbicidal activity against Arabidopsis thaliana when applied either by addition to growth media or by spraying the leaves. Transgenic Arabidopsis plants which over-express CtpA showed greater resistance to the compounds than wild-type plants providing evidence that these inhibitors are directly acting against CtpA.     

Neutral models for testing landscape hypotheses

Neutral landscape models were originally developed to test the hypothesis that human-induced fragmentation produces patterns distinctly different from those associated with random processes. Other uses for neutral models have become apparent, including the development and testing of landscape metrics to characterize landscape pattern. Although metric development proved to be significant, the focus on metrics obscured the need for iterative hypothesis testing fundamental to the advancement of the discipline. We present here an example of an alternative neutral model and hypothesis designed to relate the process of landscape change to observed landscape patterns. The methods and program, QRULE, are described and options for statistical testing outlined. The results show that human fragmentation of landscapes results in a non-random association of land-cover types that can be describe by simple statistical methods. Options for additional landscape studies are discussed and access to QRULE described in the hope that these methods will be employed to advance our understanding of the processes that affect the structure and function in human dominated landscapes.     

Expression Patterns of the Serine Protease Gene AlSP 3 in Apolygus lucorum (Hemiptera: Miridae) at Different Temperatures

As an important digestive enzyme gene, the serine protease gene AlSP 3 plays a key role in Apolygus lucorum for digesting conventional cotton. To understand the effect of different temperatures on the expression level of AlSP 3, we analyzed the expression patterns of AlSP 3 in A. lucorum reared at different temperatures using real-time polymerase chain reaction. The results showed that at the normal temperature of 24 ℃, the expression of AlSP 3 in A .lucorum at different nymphal stages was similar. The expressions of AlSP 3 in the female adult at the pre-mating stage and the male adult at post-mating stage were 19.71- and 7.16-fold higher than their control at the 1^st nymphal stage, respectively, which were significantly higher than at any other stages. However, expression in the 4^th to 5^th A. lucorum at the nymphal stage increased with rising temperature in the range of 21～30 ℃. At 30 ℃, the expression of AlSP 3 in the 2nd to 5^th A. lucorum was the highest. The expression levels of AlSP 3 in adult A. lucorum reared at different temperatures during the new emergence and pre-mating stages were significantly higher than that of the control reared at 24 ℃ (P < 0.01), while the expression during the post-mating stage at 18℃ was higher than that at other temperatures. When incubated at extreme temperatures (4 ℃ and 40 ℃), the expression of AlSP 3 in female and male adult A. lucorum was significantly higher than that at 24 ℃ (P < 0.01). Taken together, our results showed that the external environment temperature and the internal stage of A. lucorum or its sex all had significant effects on the expression of AlSP 3 in A. lucorum (P < 0.01), and that there was some interactions between them. Therefore, AlSP 3 is very important for the female adult A. lucorum to obtain nutrition at the early stages and temperature is an important limiting factor for the expression of AlSP 3.     

松香酰胺在中性造纸施胶中的应用

利用松香与异腈酸的酰经反应在１００－１５０℃下合成了三个松香酰胺，并应用它们到中性造纸松香乳酸，ＰＨ４－８的抄造试验表明这些松香衍生物在这个ＰＨ范围内具有高的施胶效果。松香酰结构与酰胺化度对施胶效果有很大的影响。