绵羊瘤胃微生物胞外蛋白酶性质的研究

本试验测定了pH、温度和EDTA对纯化的绵羊的瘤胃微生物胞外蛋白酶活性的影响,并用一已知氨基酸排列顺序的肽段(由15个氨基酸组成)作为底物,进一步测定了蛋白酶的酶切位点。结果表明,(1)纯化蛋白酶的最适pH在6.0～6.5之间,最适温度为40℃左右。(2)不同浓度的EDTA(浓度分别为1,10,25,50,75和100mM)对蛋白酶活性没有影响,因而该酶属于丝氨酸蛋白酶类。(3)纯化蛋白酶是一种内肽酶,其酶切位点在组氨酸—酪氨酸(?)、天冬氨酸—丙氨酸(?)、亮氨酸—赖氨酸(?)和(或)缬氨酸—赖氨酸(?)相连的肽键。(4)胰蛋白酶和纯化的蛋白酶虽同属丝氨酸蛋白酶类,但前者对底物具有高度的专一性,后者对底物的专一性不强,这是瘤胃内饲料蛋白质强烈降解的主要原因之     

BmNPV半胱氨酸蛋白酶基因的核苷酸序列研究

对家蚕核型多角体病毒苏州株（BmNPVsu）光胱氨酸蛋白酶基因（CP）的序列分析表明,该基因读码框为972个核苷酸,编码323个氨基酸。同源性分析表明,BmNPVsu的CP与首蓿银纹夜蛾核型多角体病毒（AcNPV）、美国白蛾核型多角体病毒（HcNPV）、云杉卷叶蛾核型多角体病毒（CfNPV）、黄杉毒蛾核型多角体病毒（OpNPV）、舞毒蛾核型多角体病毒（Ld-NPV）在DNA水平上的同源性分别为96.5％、76.2％、74.9％、72.7％、62.9％;在氨基酸水平上的同源性分别为96.9％、77.1％、79.3％、77.1％、65.6％。BmNPV CP的氨基酸序列与不同来源的木瓜蛋白酶超家族的CP也具有较高的同源性,特别与 Trpanosoma brucei的CP具有较高的一致性,达32％。在组织蛋白酶B、H、L、S以及木瓜蛋白酶的36个保守氨基酸残基中有31种出现在BmNPVsu的CP中,BmNPVsu CP同其它杆状病毒CP一样,可看作木瓜蛋白酶超家族成员。     

蝴蝶兰切花花被不同开放阶段酶活性的变化

以蝴蝶兰鲜切花为试材,对切花花被不同生理阶段3种酶活性的变化进行测量分析,以期为蝴蝶兰切花寿命的延长提供参考依据。结果表明:萼片和花瓣中的过氧化物酶(POD)从花苞期到半开期酶活性变化不明显,从半开期到衰老期酶活性显著升高。花瓣中的超氧化物歧化酶(SOD)从花苞期到半开期酶活性升高,从半开期到衰老期酶活性显著降低;萼片中的超氧化物歧化酶活性变化不显著但也呈先上升后下降趋势。花瓣中的中性蛋白酶(NP)从花苞期到半开期酶活性呈上升趋势,半开期到盛开期呈下降趋势,盛开期到衰老期呈上升趋势;花萼中的中性蛋白酶从花苞期到盛开期酶活性呈下降趋势,从盛开期到衰老期呈上升趋势;总体来看中性蛋白酶的活性呈先下降后上升趋势。     

蛹虫草CmKexin基因克隆和菌丝体中表达检测

为了探讨蛹虫草类枯草杆菌蛋白酶(Subtilisin-like protease)的表达特性,以真菌蛹虫草菌丝体为研究对象,通过RT-PCR获得蛹虫草CmKexin基因的ORF序列,并进行序列分析。应用Northern杂交和Real-time PCR方法,检测蓝光照射后蛹虫草菌丝体中CmKexin基因的转录情况。结果获得蛹虫草CmKexin基因的ORF序列。对翻译蛋白质产物CmKexin进行生物信息学分析,表明该蛋白质含1个属蛋白转化酶的肽酶S8家族功能域(143~429)和1个前体蛋白转化酶P功能域(514~600),符合真菌类枯草杆菌蛋白酶的特征。蛹虫草菌丝体在黑暗预培养4 d后再蓝光照射,Northern Blotting和Real-time PCR检测均可得到相同的结果,即在持续的蓝光照射48~50 h时,出现CmKexin基因的瞬时大量转录。而其他时间内均未检测到该基因的大量转录。试验结果将为蛹虫草类枯草杆菌蛋白酶的利用提供理论依据。     

恩施与国外白肋烟中性致香成分含量的差异比较

为明确恩施白肋烟中性致香成分组分和含量及与国外白肋烟的异同,以恩施不同产区及美国、马拉维白肋烟样品为材料,采用气质联用仪对样品的中性致香成分含量进行分析测定,结果表明:恩施白肋烟中性致香成分总量及各组分含量在产地之间存在一定差异,其中香叶基丙酮、苯甲醛、糠醇等单一致香成分含量的变异较大。以巨豆三烯酮为代表的类胡萝卜素类降解产物含量以美国烟叶最高,恩施次之,马拉维最低;恩施白肋烟苯丙氨酸类、棕色化反应产物含量均高于美国和马拉维,而西柏三烯的主要降解产物茄酮则低于国外烟叶。聚类分析将供试样品分为3类,80%的恩施样点与美国同属一类,表明恩施白肋烟中性致香成分含量与美国具有较高的相似性。     

Increased abundance of patatins,lipoxygenase and miraculins in a thaxtomin A-habituated potato Russet Burbank somaclone with enhanced resistance to common scab

Potato common scab caused by the actinobacterium Streptomyces scabiei is characterized by the formation of corky lesions on tubers that reduce their marketability. Management of common scab is very complex and often ineffective under various environmental conditions. Using potato varieties that are more resistant to common scab remains one of the most efficient strategies to control this disease. However, very little is known about the factors associated with resistance to common scab. Somaclone RB9 regenerated from thaxtomin A-habituated potato Russet Burbank calli produced tubers more resistant to common scab than the original variety. Comparison of the RB9 tuber proteome with that of Russet Burbank using label-free quantitative proteomic analysis revealed changes in the accumulation of defence-related proteins from the patatin and lipoxygenase (LOX) families, which are involved in the metabolism of lipids, and of two miraculins of the Kunitz-type protease inhibitors family. The implication of LOX during common scab infection was studied using synchronized minitubers developed from leaf-bud cuttings. S. scabiei infection stimulated the accumulation of LOX in both Russet Burbank and RB9 minitubers, but this accumulation was intensified in RB9 minitubers. Infection also increased LOX activity in Russet Burbank and RB9 minitubers. However, LOX activity measured in noninfected RB9 minitubers was similar to that of infected Russet Burbank minitubers, indicating endogenous activation of LOX activity in RB9 minitubers. We discuss how increased LOX abundance and activity in the somaclone RB9 may contribute to improving tuber defence against common scab.     

土壤铅污染对蚯蚓体腔细胞溶酶体的毒性效应

以蚯蚓溶酶体膜的稳定性作为亚细胞水平的生物标志物,研究土壤铅污染的毒性效应。将赤子爱胜蚓（Eiseniafetida）暴露于不同浓度乙酸铅染毒的人工土壤,染毒浓度为60、120、480、2400mg·kg^-1,测定蚯蚓体腔细胞溶酶体的中性红保留时间（NRRT）。结果表明,染毒6d后,480mg·kg^-1及2400mg·kg^-1剂量组,NRRT显著下降,抑制率分别为63.06%、68.61%;60mg·kg^-1及120mg·kg^-1剂量组,溶酶体膜稳定性变化不大。染毒14d后,各染毒组NRRT显著低于对照组,抑制率分别为49.43%、56.29%、68.57%、77.14%。将赤子爱胜蚓暴露于沈阳市青年公园、劳动公园、友谊宾馆、冶炼厂原址的现场土壤样品中,随着蚯蚓与土壤接触时间的增加,中性红保留时间逐渐降低,14d后各采样点蚯蚓的中性红保留时间关系为冶炼厂原址〈青年公园〈劳动公园〈友谊宾馆,与土壤铅含量的大小变化相吻合,其中冶炼厂原址差异最为显著,NRRT抑制率为92.55%,表明土壤重金属铅污染对土壤生态系统存在潜在的毒害。     

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.