温度与光照对安徽虫瘟霉产孢格局的影响*

在不同温度（10～30℃）与光照（连续光照和连续黑暗）组合条件下进行了安徽虫瘟霉（zoophthoraanhuiensis）离体产孢格局的研究。结果表明,适于安徽虫瘟霉产孢的温度为10～20℃,但以15℃最适,不仅产孢快和产孢量大,而且不受光照的影响。在25℃下虽能产孢,但产孢量大幅减少,30℃下则不产孢,25℃可能是安徽虫瘟霉产孢的高温极限。全光照各温度处理的产孢量总是比全黑暗相应温度处理的产孢量高,说明光照对产孢具有刺激作用。在10℃和全光照的组合中,累计产孢量最高且持续产孢时间最长,而相同温度与全黑暗处理的产孢量却很低,显示光照在偏低温度下是影响安徽虫瘟霉产孢的关键因素。概而言之,光照主要影响安徽虫瘟霉的产孢量,而温度主要影响其产孢速率或进程。     

温度与光照对安徽虫瘟霉产孢格局的影响

在不同温度（１０－３０℃）与光照（连续光照和连续黑暗）组合条件下进行了安徽虫瘟霉（Ｚｏｏｐｈｔｈｏｒａ ａｎｈｕｉｅｎｓｉｓ）离体产孢格局的研究。结果表明,适于安徽虫瘟霉产孢的温度为１０－２０℃,但以１５℃最适,不仅产孢快和产孢量大,而且不受光照的影响。在２５℃下虽能产孢,但产孢量大幅减少,３０℃下则不产孢,２５℃可能是安徽虫瘟霉产孢的高温极限。全光照各温度处理的产孢量总是比全黑暗相应温度处理的产     

高等植物光系统Ⅱ中强光照射产生

利用自旋捕捉电子顺磁共振(ESR)的方法对从菠菜叶绿体中分离提纯的光系统Ⅱ(PSⅡ)颗粒产生Q2的机理进行了直接检测.通过对样品充氧、加入超氧化物歧化酶(SOD)抑制剂四氰乙烯(TCNE)以及原位光照检测ESR信号等手段,在PSⅡ中检测到O2与DMPO加合物的特征ESR信号.而在没有SOD抑制剂的情况下,光照时PSⅡ中O2与DMPO加合物浓度显著下降.进一步实验发现PSⅡ中O2产率与氧分子浓度直接正相关.O2产率还具有pH值依赖性,在pH值为6.0～6.5范围内,O2产率最高,大于此范围时则呈显著下降趋势.而PSⅡ颗粒的Tris处理也将导致O2产率的急剧减少.以上结果证实水裂解放氧十分活跃的PSⅡ也是高等植物叶绿体在光照下产生活性O2的主要部位,通常大部分的O2能被内源SOD清除,且O2的生成与PSⅡ的电子传递活性密切相关.     

光周期对飞虱虫疠霉离体培养的生物量及其产孢节律的影响

飞虱虫疠霉（Ｐａｎｄｏｒａｄｅｌｐｈａｃｉｓ）是侵染飞虱和叶蝉等重要作物害虫的昆虫病原真菌。通过在液体及平板培养中研究光周期对菌丝及菌落生长量、产孢节律和产孢量的影响,作者发现光照对液培菌丝的营养生长无显著影响,但长光照液培菌丝的产孢延迟并且产孢量减少,而全黑暗液培菌丝产孢早且产孢量大。但是,光照明显促进全黑暗下液培菌丝的产孢。在平板培养中,长光照能促进菌落生长和产孢;半光照半黑暗虽有利菌落生长,但产孢量很低;光照９ｈ和１８ｈ产孢量较大,但菌落生长较小。无论液体还是平板培养,飞虱虫疠霉均生长良好,但建议在全黑暗条件下进行液体培养,在长光照下进行平板培养。     

高等植物光系统Ⅱ中强光照射产生地超氧阴离子自由基的ESR探索

利用自旋捕捉电子顺磁共振(ESR)的方法对从菠菜叶绿体中分离提纯的光系统Ⅱ(PSⅡ)颗粒产生Q2的机理进行了直接检测.通过对样品充氧、加入超氧化物歧化酶(SOD)抑制剂四氰乙烯(TCNE)以及原位光照检测ESR信号等手段,在PSⅡ中检测到O2与DMPO加合物的特征ESR信号.而在没有SOD抑制剂的情况下,光照时PSⅡ中O2与DMPO加合物浓度显著下降.进一步实验发现PSⅡ中O2产率与氧分子浓度直接正相关.O2产率还具有pH值依赖性,在pH值为6.0～6.5范围内,O2产率最高,大于此范围时则呈显著下降趋势.而PSⅡ颗粒的Tris处理也将导致O2产率的急剧减少.以上结果证实水裂解放氧十分活跃的PSⅡ也是高等植物叶绿体在光照下产生活性O2的主要部位,通常大部分的O2能被内源SOD清除,且O2的生成与PSⅡ的电子传递活性密切相关.     

转基因莱茵衣藻hemHc-lbac 和根瘤菌共培养提高产氢培养条件的优化

转基因莱茵衣藻hemHc-lbac(transgenic Chlamydomonas reinhardtii hemHc-lbac)以不同比例与日本慢生大豆根瘤菌(Bradyrhizobium japonicum)混合, 在不同光照条件下进行产氢培养, 以确定产氢的最优条件和探索产氢提高的机理。结果表明藻菌共培养的最优产氢条件为25 ℃、光照30 μE⋅m–2⋅s–1、生长至饱和期的菌和藻体积比为1: 80, 产氢量达到最大, 约为278 μmol⋅mg–1Chl, 是对照组80 μmol⋅mg–1Chl 的3.5 倍。藻菌共培养提高产氢量的主要原因是体系中氧气浓度的降低而使氢化酶活性提高、以及衣藻生物量的增加。该研究为利用藻菌共培养及转基因的方法提高微藻光合生物制氢效率提供了重要实验基础。     

两种典型养鸡模式的能值分析

畜禽养殖的有机化过程被认为是实现养殖业持续发展与保障食品安全的重要途径.运用能值分析方法对两种典型养鸡模式——家庭有机饲养和林地散养进行了系统分析,并结合Castellini等人对意大利所做的同类研究(工业规模化笼养和草地散养)进行了横向对比.结果表明,林地散养生产系统的能值产出率为1.11,与家庭有机饲养系统相近,而在环境负载率和可持续发展能力方面的表现,均差于家庭有机饲养生产系统,说明林地散养模式尽管通过提高系统生产空间,理论上接近畜禽养殖的有机化理念,但由于大量的禽舍建筑、医药及其饲料等方面的外部资源输入,反而使得系统的环境负载率提高,可持续性大大下降.与意大利的结果比较发现,我国两个养殖系统的社会经济成本高、对外部不可再生资源的依赖程度较高,有机化目标并没有很好的实现.因此,我国现有的有机化畜禽养殖方式和外部制度都有待进一步改进和创新.     

深红红螺菌吸氢酶缺失突变株在管式光合反应器中的产氢

本研究设计了一种2 L分体式管式光合反应器, 并研究了深红红螺菌(Rhodospirillum rubrum)吸氢酶缺失突变株在该反应器中分别利用人工光源(持续光照与光暗交替)和自然光的产氢规律。结果表明在人工光照条件下R. rubrum的产氢可维持5 d, 持续光照和光暗交替条件下(12 h: 12 h)的氢产量可分别达到5752 mL/PBR ± 158 mL/PBR和5012 mL/PBR ± 202 mL/PBR; 自然光条件下, 最适产氢光照强度为30000 Lux~40000 Lux; 在此光照条件下, R. rubrum产氢可维持6 d~ 10 d, 最高氢产量可达到2800 mL/PBR。尽管利用自然光的氢产量比利用人工光源氢产量低, 但是利用自然光的产氢比较经济, 并且该光合产氢系统操作简单, 该工艺有望开发为低成本的光合细菌产氢技术。     

光照和温度对望天树种子萌发的影响

在实验室内人工气候箱控制的条件下,研究了我国重要珍稀濒危植物望天树（Shorea wantianshuea,龙脑香科（Dipterocarpaceae））种子萌发对持续光照、14小时光照／10小时黑暗周期性光照的反应;同时研究了望天树种子的萌发对不同温度的反应和低温贮藏对种子活力的影响。结果表明,不论是持续光照还是周期性光照都不能提高望天树种子的萌发率,相反,持续光照和周期性光照都不同程度降低了种子萌发率。光照能通过加速或延迟种子萌发的进程、或改变幼苗活力指数和萌发指数而影响种子萌发的质量;持续光照延迟种子萌发的进程,而周期性光照加快种子萌发的进程。30℃是种子的最适萌发温度,虽然15℃和5℃的相对低温对幼苗活力指数影响不大,但大大延迟了种子萌发进程,并提高萌发率。望天树种子不能耐受5℃低温贮藏,但具有在15℃下短期贮藏的潜力和一定程度的生理性休眠。     

光照和温度对望天树种子萌发的影响

在实验室内人工气候箱控制的条件下, 研究了我国重要珍稀濒危植物望天树 (Shorea wantianshuea,龙脑香科 (Dipterocarpaceae)) 种子萌发对持续光照、14小时光照 / 10小时黑暗周期性光照的反应; 同时研究了望天树种子的萌发对不同温度的反应和低温贮藏对种子活力的影响。结果表明, 不论是持续光照还是周期性光照都不能提高望天树种子的萌发率, 相反, 持续光照和周期性光照都不同程度降低了种子萌发率。光照能通过加速或延迟种子萌发的进程、或改变幼苗活力指数和萌发指数而影响种子萌发的质量; 持续光照延迟种子萌发的进程, 而周期性光照加快种子萌发的进程。30℃是种子的最适萌发温度, 虽然15℃和5℃的相对低温对幼苗活力指数影响不大, 但大大延迟了种子萌发进程, 并提高萌发率。望天树种子不能耐受5℃低温贮藏, 但具有在15℃下短期贮藏的潜力和一定程度的生理性休眠。     

Production of bioactive lysophosphatidic acid by lysophospholipase D in hen egg white

Lysophosphatidic acid (LPA), a lysophospholipid mediator, is produced extracellularly by lysophospholipase D (lysoPLD) secreted in several animal body fluids including blood plasma. Previously, we reported that hen egg white contains polyunsaturated fatty acid-rich LPA. In this study, we examined whether lysoPLD is involved in the production of LPA in hen egg white. LysoPLD activity was measured by determining LPA and choline by mass spectrometric and enzyme-linked fluorometric analyses, respectively. LysoPLD increased with increased dilution of egg white, indicating that one or more components of egg white strongly inhibit its lysoPLD activity. This dilution-dependent increase in the lysoPLD activity was masked by co-incubation of the egg white with lysozyme, a major protein in hen egg white. Furthermore, addition of Zn(2+), Mn(2+), Ni(2+), or Co(2+) to diluted egg white altered preference patterns of lysoPLD toward choline-containing substrates. In particular, the egg white lysoPLD activity was greatly increased when Co(2+) was added. The cation-requirement of lysoPLD activity in hen egg white resembled that of plasma autotaxin (ATX)/lysoPLD. Western blot analysis revealed that egg white contained a protein that was immunostained with anti-ATX antibody. These results suggested that LPA in hen egg white is produced from lysophospholipids, especially LPC, by the action of ATX/lysoPLD, possibly originating from hen oviduct fluid.     

Persistency of broodiness in recycled turkey hens

Large White turkey breeder hens identified as broody during their initial egg production cycle were recycled into a second and third reproductive cycle in order to determine the persistency of broodiness. Egg production data included time to first egg, duration of lay and the number of eggs per hen. Nesting data included time to first broody nesting activity and duration of broody nesting activity. In general, there were no significant differences among the three cycles for the above observations. Mean values over the three cucles were as follows: 24.6 days to first egg following photostimulation; 49.4 days of egg production; 29.4 eggs per hen; 46.9 days from the first egg to first broody nesting and 59.7 days duration of nesting. Of 10 hens identified as broody in their first egg production cycle and followed through two additional 20-week cycles, seven hens demonstrated broody nesting behavior and nine had low rates of egg production. It was concluded that broody traits of a given hen are carried over into subsequent cycles.     

Involvement of osteopontin in egg shell formation in the laying chicken

Expression of the osteopontin (OPN) gene in the oviduct of the laying hen was studied. It was detected only in the egg shell gland (ESG), where massive calcification occurs. No OPN gene expression was detected in any other part of the oviduct, such as the magnum and isthmus. The OPN gene was expressed in a circadian fashion during the daily egg cycle only during the period of egg shell calcification. No OPN gene expression was detected in the ESG of a pre-laying hen before the onset of reproduction, or after forced removal of the egg close to its entrance into the ESG. OPN was found to be synthesized by the epithelial cells of the ESG lining the lumen. Upon synthesis, OPN is immediately secreted out of cells and accumulates in the egg shell. These findings demonstrate for the first time temporal and spatial association of OPN with egg shell calcification. OPN, which was found to be part of the organic matrix of the egg shell, may play an important role in egg shell calcification.     

Amyloid protofilament formation of hen egg lysozyme in highly concentrated ethanol solution

Mutant human lysozymes (Ile56Thr & Asp67His) have been reported to form amyloid deposits in the viscera. From the standpoint of understanding the mechanism of amyloid formation, we searched for conditions of amyloid formation in vitro using hen egg lysozyme, which has been extensively studied from a physicochemical standpoint. It was found that the circular dichroism spectra in the far-ultraviolet region of the hen egg lysozyme changed to those characteristic of a beta-structure from the native alpha-helix rich spectrum in 90% ethanol solution. When the concentration of protein was increased to 10 mg/mL, the protein solution formed a gel in the presence of 90% ethanol, and precipitated on further addition of 10 mM NaCl. The precipitates were examined by electron microscopy, their ability to bind Congo red, and X-ray diffraction to determine whether amyloid fibrils were formed in the precipitates. Electron micrographs displayed unbranched protofilament with a diameter of approximately 70 A. The peak point of the difference spectrum for the Congo red binding assay was 541 nm, which is characteristic of amyloid fibrils. The X-ray diffraction pattern showed a sharp and intense diffraction ring at 4.7 A, a reflection that arises from the interstrand spacing in beta-sheets. These results indicate that the precipitates of hen egg lysozyme are amyloid protofilament, and that the amyloid protofilament formation of hen egg lysozyme closely follows upon the destruction of the helical and tertiary structures.     

Infradian rhythmicity in egg production features in relation to antioxidant profiles of Rhode Island Red (RIR) birds reared at backyard in different agroclimatic zones of West Bengal during summer stress

Egg production features viz. weekly hen day and egg weight together with some stress markers were studied in RIR birds reared under backyard in different agroclimatic zones of West Bengal. Overall, weekly hen day average and egg weight in summer was 3.39 ± 0.09 and 45.13 ± 0.24 g, respectively. But the pattern of egg production in various zones is not same, as significant egg production difference (P ≤ 0.01) among zones was noticed in summer, from 26th to 37th week. There was significant (P ≤ 0.01) correlation between weekly hen day average and egg weight on 26th, 35th and 37th week (P ≤ 0.05). The observed overall level of antioxidant enzymes viz. SOD (U/g of Hb), glutathione peroxidase (GSH-Px) (μmol/L), TAS (mmol/L) and LDH (IU/L) were 0.56 ± 0.61, 565.15 ± 0.61, 1.43 ± 0.61 and 203.05 ± 0.61, respectively, irrespective of zones. It was observed that mean concentration of SOD, GHS-Px, TAS and LDH of RIR birds reared at backyard have positive association with weekly hen day average and average egg weight throughout summer stress. The current findings showed that RIR birds reared at backyard had better adaptation ability to summer stress in different agroclimatic zones of West Bengal.     

Detection of chitinase activity after polyacrylamide gel electrophoresis

Commercial Streptomyces griseus and Serratia marcescens chitinases and purified wheat germ W1A and hen egg white lysozymes were subjected to polyacrylamide gel electrophoresis under native conditions at pH 4.3. After electrophoresis, an overlay gel containing 0.01% (W/V) glycol chitin as substrate was incubated in contact with the separation gel. Lytic zones were revealed by uv illumination with a transilluminator after staining for 5 min with 0.01% (W/V) Calcofluor white M2R. As low as 500 ng of purified hen egg lysozyme could be detected after 1 h incubation at 37 degrees C. One band was observed with W1A lysozyme and several bands with the commercial microbial chitinases. The same system was also used with native polyacrylamide gel electrophoresis at pH 8.9. Several bands were detected with the microbial chitinases. The same enzymes were also subjected to denaturing polyacrylamide gel electrophoresis in gradient gels containing 0.01% (W/V) glycol chitin. After electrophoresis, enzymes were renatured in buffered 1% (V/V) purified Triton X-100. Lytic zones were revealed by uv after staining with Calcofluor white M2R as for native gels. The molecular weights of chitinolytic enzymes could thus be directly estimated. In denaturing gels, as low as 10 ng of purified hen egg white lysozyme could be detected after 2 h incubation at 37 degrees C. Estimated molecular weights of St. griseus and Se. marcescens were between 24,000 and 72,000 and between 40,500 and 73,000, respectively. Some microbial chitinases were only resistant to denaturation with sodium dodecyl sulfate while others were resistant to sodium dodecyl sulfate and beta-mercaptoethanol.     

Structural characteristics of hen egg ovalbumin expressed in yeast Pichia pastoris

The recombinant ovalbumin (OVA) produced in yeast Pichia pastoris was purified from the culture medium by anion exchange chromatography, and its structural characteristics were compared with those of hen egg OVA, mainly from the point of view of posttranslational modification. The expressed OVA consisted of two molecular species immmunoreactive with antibody for hen egg OVA. The two molecular species, 45 and 47 kDa in molecular size, were thought to correspond to mono-glycosylated form and di-glycosylated form respectively. The non-glycosylated form was not produced in the system. The other posttranslational modifications (N-terminal acetylation and phosphorylation) observed in hen egg OVA were not detected in either of the molecular species. The two recombinant proteins displayed almost exactly the same circular dichroism and intrinsic tryptophan fluorescence spectra as hen egg OVA. The melting temperature, Tm, which was determined from the thermal unfolding curve, was almost identical in the two recombinant proteins, despite the difference in glycosylation levels, while it decreased by about 2.5 degrees C as compared with that of hen egg OVA (77.3 degrees C). These data indicate that the additional glycosylation to Asn-311 in the recombinant protein does not affect protein conformation or thermostability.     

Nutrient and drug effects on cholesterol metabolism in the laying hen

The laying hen is a highly dynamic model for studies of cholesterol metabolism. Cholesterol biosynthesis takes place primarily in the liver where it is regulated by both diet and drugs. Ovarian cholesterol biosynthesis follows a pattern different from that in liver and is not influenced by dietary fat or cholesterol. The hen responds to high levels of dietary polyunsaturated fat by increasing cholesterol biosynthesis, egg cholesterol deposition, and fecal bile acid excretion. Dietary cholesterol curtails liver cholesterol biosynthesis and may or may not result in increased egg cholesterol deposition and/or increased fecal steroid excretion depending on the level of cholesterol intake. Dietary plant sterols and fiber may moderate egg cholesterol deposition but the conditions under which this takes place are not well defined. D-Thyroxin reduces blood cholesterol, increases blood sterol turnover, and increases egg cholesterol concentration. Triparanol and azasterols prevent desmosterol conversion to cholesterol with resultant appearance of both sterols in blood and eggs. Probucol moderates egg cholesterol deposition by reducing synthesis and/or transfer of the sterol to the egg. Implications for the use of the hen in cholesterol metabolism studies are discussed.     

Comparative Studies of Yolkin Preparations Isolated from Egg Yolks of Selected Bird Species

The results of our research have proven that yolkin preparations isolated from eggs of different bird species show a high similarity in polypeptide composition. Despite the small differences in protein patterns, all of yolkin preparations showed also strong immunomodulatory activity, comparable with yolkin obtained previously from hen egg yolk. It can therefore be deducted that the presence of this polypeptide complex in the egg is not accidental and performs an important biological function for developing embryo.     

Egg Yolk antibody and its application

A hen transfers her serum immumnoglobulin G to the egg yolk (IgY) and gives im|munity to her offspring. Therefore, the hen egg can be an effective supplier of a large amount of antigen specific antibody that accumulates in the egg yolk. Antigen specific antibody has been widely used for immunological analysis in the field of diagnosis as well as pure scientific research. The production and separation technology of IgY is demonstrated in the present study.