外源NO及反向调控PEG胁迫下苜蓿萌发种子抗氧化酶及其同工酶动态的研究

以紫花苜蓿为材料,用一氧化氮供体硝普钠(SNP)及一氧化氮清除剂c-PTIO对苜蓿种子进行浸种处理,采用分光光度法和同工酶电泳技术来研究外源NO及反向调控对PEG胁迫下紫花苜蓿抗氧化酶活性及其同工酶的影响,并探讨NO调控苜蓿种子耐旱性的生理机制。结果表明:PEG胁迫下外施0.1 mmol·L^-1 SNP,第6天时较PEG处理POD活性降低了32.72%;第4天时SOD、CAT活性较PEG处理升高了10.48%、23.60%,有效缓解了PEG对紫花苜蓿萌发中种子的氧化损伤,提高其抗氧化能力。PEG胁迫下添加 c-PTIO抑制了苜蓿萌发期的抗氧化系统活性。从同工酶的谱带数量和强弱来看,POD同工酶各区带活力均随PEG胁迫时间的延长而增加,在第2天酶带只有1条,而第4天酶带呈现9条;SOD和CAT同工酶表达量变化不显著,但酶带强弱有一定变化,S3酶带随处理时间的延长表达量逐渐减弱;CAT同工酶谱带则一直保持2条带,无明显强弱变化。因此,外源NO在PEG胁迫下紫花苜蓿萌发中抗氧化酶快速响应并在维护氧自由基代谢平衡中起重要保护作用。     

Colonization and histopathology of susceptible and resistant carnation cultivars infected with Fusarium oxysporum f. sp. dianthi

Stems of the susceptible Early Sam and resistant Novada carnations were inoculated with a conidial suspension ofFusarium oxysporum f. sp.dianthi. Stem segments of either cultivar were sampled regularly and used for determination of fungal growth and for microscopical investigation.Early Sam showed typicalFusarium wilt symptoms and its stems were colonized intensively. The observed vascular browning appeared to be caused by discolouration of primary walls of infected vessels and surrounding cells. Vessels were rarely occluded with gel. Cell wall degradation led to the formation of stem cavities. Hyperplasia of xylem parenchyma was not seen.In Novada, fungal colonization remained low throughout the experiment. Macroscopic symptoms were absent except for longitudinal bursts in the stem, which appeared to be caused by hyperplasia of xylem parenchyma bordering infection. Vascular gelation occurred in the infected tissues, causing some vascular browning also. Xylem vessel regeneration was observed in the hyperplastic layer. Cavities were not formed, and wall discolouration was rare. Vascular gelation is considered part of theFusarium wilt resistance mechanism. It is followed by xylem vessel regeneration, which expresses a general plant response to vascular dysfunction rather than being part of the resistance mechanism.Although of different origin, vascular browning as such occurs in both susceptible and resistant interactions. In breeding for resistance, care should hence be taken with the current use of browning as an indication of disease.Samenvatting Anjers van de vatbare cultivar Early Sam en de resistente cultivar Novada werden geïnoculeerd met een conidiënsuspensie vanFusarium oxysporum f. sp.dianthi. Van beide cultivars werden regelmatig stengeldelen geoogst om deze microscopisch te onderzoeken en om de schimmelgroei te bepalen.Early Sam vertoonde de voor deze verwelkingsziekte kenmerkende symptomen en werd intensief gekoloniseerd. Aan het vaatweefsel waargenomen bruinkleuring bleek veroorzaakt te worden door verkleuring van de primaire wanden van geïnfecteerde vaten en de hen omringende cellen. Zelden trad er in de vaten gomvorming op. Celwandafbraak veroorzaakte de vorming van holten in de stengel. Hyperplasie van het houtparenchym werd niet waargenomen.In Novada bleef de schimmelgroei gedurende het hele experiment beperkt. Macroscopisch waren er enkel lengtescheuren in de stengel te zien, die veroorzaakt bleken te worden door hyperplasie van aan de infectie grenzend houtparenchym. In het geïnfecteerde vaatweefsel optredende gomvorming veroorzaakte ook enige bruinkleuring. In het hyperplastische weefsel werd regeneratie van houtvaten waargenomen. In de stengel werden geen holten gevormd, en verkleuring van de celwanden kwam weinig voor. De vorming van gommen in de houtvaten maakt waarschijnlijk deel uit van het resistentiemechanisme. De daarop volgende houtvatregeneratie is eerder een algemene reactie van de plant op vaatverstopping dan een deel van het resistentiemechanisme.Vaatverbruining, zij het van verschillende oorsprong, komt voor in zowel vatbare als resistente interacties. Om die reden moet men in de resistentieveredeling bij de anjer voorzichtig zijn met het gebruik van bruinkleuring als ziekteïndicatie.     

Lipoprotein electrophoresis differentiation of chylous and nonchylous pleural effusions in dogs and cats and its correlation with pleural effusion triglyceride concentration

Pleural effusions from 23 dogs and 25 cats hospitalized between November 1986 and April 1987 at the Veterinary Hospital of the University of Pennsylvania (VHUP) were classified as chylous (chylomicrons present) or nonchylous (chylomicrons absent) by the presence or absence, respectively, of a chylomicron band on lipoprotein electrophoresis gels of the effusions. Triglyceride concentrations, cholesterol concentrations, and cholesterol/triglyceride (C/T) ratios were compared between the chylous and nonchylous groups in each species. Cholesterol concentrations were not significantly different between chylous and nonchylous effusions in both dogs and cats. Cholesterol/triglyceride ratios of less than 1 were present in all dogs and cats with chylous effusions; however, 12% of dogs with nonchylous effusions and 50% of cats with nonchylous effusions also had a C/T ratio less than 1. Triglyceride concentrations accurately classified all effusions as chylous or nonchylous in both dogs and cats. Pleural effusions with triglyceride concentrations greater than l00 mg/dl were chylous in all cases, whereas, effusions with triglyceride concentrations less than l00 mg/dl were nonchylous in all cases.     

鹰嘴豆籽粒贮藏蛋白双向电泳的研究

以鹰嘴豆为材料,通过对样品制备、电泳条件的优化,建立其籽粒贮藏蛋白的双向电泳技术,并通过电泳图谱的分析,了解贮藏蛋白等电点和相对分子质量的分布情况．结果表明,经考马斯亮蓝染色获得贮藏蛋白斑点约380个,等电点范围在pH4．0～8．0,其中大部分集中在pH6．0～7．0;鹰嘴豆籽粒贮藏蛋白质的相对分子质量为11000．120000,表现出2个分布峰,一个在30000～40000,另一个在50000～60000;鹰嘴豆与大豆籽粒贮藏蛋白的分子质量和等电点分布基本相似．     

高效中华绒蟹微卫星文库的构建及其七个位点的特征分析

一次富集回收产物与生物素标记探针杂交,运用磁珠亲和捕捉构建中华绒螯蟹微卫星文库。96个阳性克隆双向测序结果为75个克隆含有微卫星,登录号为DQ388769~DQ388807,表明二次富集法是一种快速高效的分离微卫星的方法。7个微卫星位点的特征分析显示,ES10位点是复制位点;ES37可能有空位点;其它5个位点可用于中华绒螯蟹的品种鉴定、遗传多样性和群体结构研究。     

番茄青枯病抗性两个相关基因功能的初步鉴定

对抗性番茄材料‘Hawaii 7996’分别接种强致病力青枯菌（RsH）和中等致病力青枯菌（RsM）后,利用双向电泳技术找到了两个差异表达蛋白质--谷胱甘肽S转移酶L3和Remorin 1蛋白。在利用携带八氢番茄红素脱氢酶基因PDS的TRV重组病毒载体通过叶片浸润法证明VIGS体系可行后,进一步利用该技术沉默上述两个蛋白质的对应基因GST-L3和Rem-1,并用半定量RT-PCR检测沉默效果。结果表明,分别沉默GST-L3和Rem-1的番茄植株抗性减弱,在接种中等致病力青枯菌（RsM）后青枯病发病率和病情指数都明显高于对照植株,说明基因GST-L3和Rem-1与番茄对青枯病的抗性相关。     

茭白黑粉菌脉冲电泳分析方法

采用脉冲电泳法对茭白黑粉菌染色体进行核型分析。分析结果表明,茭白黑粉菌脉冲电泳样品处理方法包括原生质体的制备、电泳参数的设置等,对试验结果有一定影响,在制备电泳样品时,包埋的原生质体或孢子的浓度不能太低,蛋白酶K的浓度不能过高,低熔点琼脂糖的浓度不能太低,此外还应确保电泳槽水平,并且电泳结束后及时取出凝胶。     

冷刺激对两种泥鳅血液蛋白质的影响

采用SDS-聚丙稀酰胺凝胶电泳技术,探讨冷刺激对泥鳅血液蛋白质的影响.结果表明,泥鳅和大鳞副泥鳅的血液蛋白质发生了变化,即产生了冷激蛋白;同种泥鳅,控制产生冷激蛋白的特定基因在一定的刺激强度和刺激程度下,产生冷激蛋白的时间和种类不同;不同种泥鳅,在相同低温处理条件下,产生冷激蛋白的时间和种类也不一样.本研究不仅为研究冷激蛋白提供了前提,而且为两种泥鳅的人工养殖提供了参考.     

马铃薯基乳液作为脂肪替代物对鱼糜凝胶品质影响研究

为了研究马铃薯基乳液（potato based emulsion, PE）对鱼糜凝胶品质的影响, 本研究将玉米油（corn oil,CO）或PE按不同添加量（0%~14%）与鱼糜混合,以鱼糜凝胶质构、水分、色泽、分子间相互作用、流变特性和微观结构为考察指标,探究PE作为脂肪替代物对鱼糜凝胶品质的影响及机理。 结果显示,在相同的油添加量下,与CO相比,PE能避免油滴对鱼糜凝胶质构特性和持水能力产生的劣化效果,使样品表现出更高的弹性模量和粘度,其中,6%油含量PE样品的硬度从1 073 g（0%油含量样品）提升到1 159 g,而6%油含量CO样品的硬度降低至837 g;另外,横向弛豫时间结果表明,PE抑制了不易流动水向由水的转变,使更多的不易流动水被束缚,这可能增强了光线漫反射,从而使PE样品白度更高;微观结构和分子间作用力测定结果表明PE样品中油滴分布更均匀,氢键、疏水相互作用和二硫交联更强,凝胶网络因淀粉颗粒的填充效应而更致密;十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果表明 ,PE组中油滴对鱼糜蛋白交联的干预程度更小。研究表明,PE可作为脂肪替代物提升鱼糜凝胶制品品质。本研究可为淀粉基脂肪替代物的开发及其在鱼糜凝胶产品中的应用提供理论基础。     

恩施碎米荠种子蛋白质双向电泳初探

对湖北恩施高硒地区生长的具有超富硒能力的植物恩施碎米荠的种子蛋白质用TCA-丙酮沉淀提取,等电聚焦采用固相梯度胶条,双向电泳分离蛋白质.结果表明,蛋白质样品制备的质量高;银染胶图背景干净,蛋白点清晰,且呈现较好的聚焦结果,无明显拖尾或横纹现象;蛋白质集中分布在pH5～10的范围之内,分子量主要在20～60 kμ的范围之内.