Improving web server performance by network aware data buffering and caching

In this paper we propose a new method of data handling for web servers. We call this method Network Aware Buffering and Caching (NABC for short). NABC facilitates reduction of data copies in web server's data sending path, by doing three things: (1) Layout the data in main memory in a way that protocol processing can be done without data copies (2) Keep a unified cache of data in kernel and ensure safe access to it by various processes and kernel and (3) Pass only the necessary meta data between processes so that bulk data handling time spent during IPC can be reduced. We realize NABC by implementing a set of system calls and an user library. The end product of the implementation is a set of APIs specifically designed for use by the web servers. We port an in house web server called SWEET, to NABC APIs and evaluate performance using a range of workloads both simulated and real. The results show a very impressive gain of 12% to 21% in throughput for static file serving and 1.6 to 4 times gain in throughput for lightweight dynamic content serving for a server using NABC APIs over the one using UNIX APIs.

Process Development for Functionalization of Cotton with Silver Nanoparticles Synthesized by Bio-based Approaches

Cotton is a widely used raw material for textiles but drawbacks regarding their poor mechanical properties often limit their applications as functional materials. The present investigation involved process development for one step coating of cotton with silver nanoparticles (SNP) synthesized using Azadirachta indica and Citrus limon extract to develop functional textiles. Addition of starch to functional textiles led to efficient binding of nanoparticles to fabric and led to drastic decrease in release of silver from fabricated textiles after ten washing cycles enhancing their environment friendliness. Differential scanning calorimetry, scanning electron microscopy, FT-IR analysis and mechanical studies demonstrated efficient binding of nanoparticles to fabric through bio-based processes. The functionalized textiles developed by the bio-based methods showed significant antibacterial activity against E. coli and S. aureus (with 99% microbial reduction). Present work offers a simple procedure for coating SNP using bio-based approaches with promising applications in specialized functions.

SWEETENING OF BIOGAS IN SPRAY TOWERS FOR POWER GENERATION

This paper presents the experience of the new design of using impinging jet spray columns for scrubbing hydrogen sulfide from biogas that has been developed by Indian Institute of Science and patented. The process uses a chelated polyvalent metal ion which oxidizes the hydrogen sulfide to sulfur as a precipitate. The sulfur generated is filtered and the scrubbing liquid recycled after oxidation. The process involves in bringing contact the sour gas with chelated liquid in the spray columns where H2S reacts with chelated Fe3+ and precipitates as sulfur, whereas Fe3+ gets reduced to Fe2+. Fe2+ is regenerated to Fe3+ by reaction of oxygen in air in a separate packed column. The regenerated liquid is recirculated. Sulfur is filtered and separated as a byproduct. The paper presents the experience in using the spray towers for hydrogen sulfide removal and further use of the clean gas for generating power using gas engines. The maximum allowable limit of H2S for the gas engine is 200 ppm (v/v) in order to prevent any corrosion of engine parts and fouling of the lubricating oil. With the current ISET process, the hydrogen sulfide from the biogas is cleaned to less than 100 ppm (v/v) and the sweet gas is used for power generation. The system is designed for 550 NM3/hr of biogas and inlet H2S concentration of 2.5 %. The inlet concentration of the H2S is about 1 - 1.5 % and average measured outlet concentration is about 30 ppm, with an average gas flow of about 300 - 350 NM3/hr, which is the current gas production rate. The sweet gas is used for power generation in a 1.2 MWe V 12 engine. The average power generation is about 650 - 750 kWe, which is the captive load of the industry. The plant is a CHP (combined heat power) unit with heat from the cylinder cooling and flue being recovered for hot water and steam generation respectively. The specific fuel consumption is 2.29 kWh/m(3) of gas. The system has been in operation for more than 13,000 hours in last one year in the industry. About 8.4 million units of electricity has been generated scrubbing about 2.1 million m3 of gas. Performance of the scrubber and the engine is discussed at daily performance level and also the overall performance with an environment sustenance by precipitating over 27 tons of sulfur.

Estudio de cinco leguminosas en la fertilización y métodos de manejo de malezas en el cultivo de la Pitahaya (Hylocereus undatus Britt y Rose)

En el mes de junio de 1996, se estableció un experimento de campo en la empresa Frutas Tropicales Ltda, localizada en la comarca de Guanacastillo, departamento de Masaya, con el propósito de evaluar la influencia de cinco leguminosas de cobertura asociadas con el cultivo de la pitahaya (Hylocereus undatus Britt & Rose), como manejo de las malas hierbas y aporte de nutrientes a partir de la materia orgánica incorporada. Al estudio se le aplicó el diseño de bloques completos al azar con cuatro repeticiones.Los tratamientos evaluados fueron: Vigna radiata (L.) Wilczek, Mucuna pruriens (L.) DC., Vigna unguiculata (L.) Walp., Canavalia ensiformis (L.) DC., Lablab purpureus (L.) Sweet y el tratamiento Testigo, el cual representa el manejo tradicional en el control de las malezas de parte del productor. Los resultados indican que el mayor peso seco acumulado lo presentó C. ensiformis al registrar 7 401 kg/ha. En tanto, V. radiata con 2 331.66 kg/ha presentó el menor valor de peso seco. En el comportamiento del peso seco de las malezas se registró mas alto en M. pruriens con 1 600 kg/ha. En tanto, L. purpureus con 545 kg/ha fue el más bajo.Para medir el aporte de nutrientes a partir de la materia orgánica se hizo un análisis de suelo previo a la siembra de las leguminosas. Un segundo y tercer análisis de suelo se efectuaron 3 meses y 11 meses después de realizado el primer análisis. En nitrógeno el mejor resultado se obtuvo en V. unguiculata al ubicarse como mejor aportador en el primer análisis con 149.60 kg/ha y segundo mejor aportador en el segundo análisis con 113.28 kg/ha.Superado levemente en este caso por el tratamiento testigo con 113.40 kg/ha. En ambos análisis el menor aportador de nitrógeno fue C. ensiformis con 122.40 kg/ha y 103.32 kg/ha para el primer y segundo análisis respectivamente.En cuanto a fósforo, V. radiata se ubicó como mejor aportador en el primer análisis con 16.80 kg/ha, mientras que en el segundo análisis se ubicó como tercero con 26.49 kg/ha, superado por C. ensiformis y V. unguiculata con 40.66 kg/ha y 31.32 kg/ha respectivamente. En cambio M. pruriens se ubicó como el aportador más bajo de fósforo con 6.60 kg/ha en el primer análisis y 3.98 kg/ha en el segundo.En relación al potasio, el mejor resultado se obtuvo en M. pruriens con 567.73 kg/ha y 694.42 hg/ha en el primer y segundo muestreo respectivamente. Mientras que L. purpureus fue el menor aportador con 384.74 kg/ha y 273.70 kg/ha en el primer y segundo muestreo respectivamente.Con respecto a la influencia sobre la dinámica de las malezas el mejor resultado se presentó en M. pruriens, al eliminar por completo la presencia de malezas a los 95 días de establecido el ensayo. Similar comportamiento tuvo C. ensiformis con la diferencia que fue en un período más prolongado. Mayor abundancia de malezas presentaron los tratamiento L. purpureus, V. radiata y V. unguiculata, en los cuales hubo necesidad de acompañarlos con controles mecánicos. Por último, se ubicó el tratamiento Testigo el cual siempre necesito de control mecánico. En relación al mayor número de brotes por planta, los tratamientos implementados no tuvieron diferencias significativas entre ellos, pero en términos numéricos los que presentaron mayor número de brotes fueron V. radiata y Testigo con 4,856 y 4,417 respectivamente, seguidos de V. unguiculata y C. ensiformis con 3,933 y 3,922 respectivamente. Por último se ubicaron M. pruriens y L. purpureus con 3,267 y 3,034 respetivamente.De acuerdo a los resultados obtenidos en cobertura de suelo, diversidad y abundancia de malezas, biomasa y aporte de nutrientes a partir de residuos vegetales, excepto en fósforo, se recomienda el uso de la leguminosa Mucuna pruriens (L.) DC. En asocio con el cultivo de la pitahaya.

Determinación de la relación sólidos solubles/acidez total en muestras de frutos de cuatro especies de cítricos

Este trabajo fue realizado en el laboratorio químico de la Estación Experimental Agropecuaria La Calera en la terminación del año de 1964 y principios de 1965 con las especies: naraja dulce (Citrus sinensis); grape fruit (Citrus paradisi); mandarina (Citrus reticulata) y limón agrio (Citrus limonum). El objeto del trabajo fue el investigar la relación sólidos totales solubles sobre acidez total en base a un tamaño promedio del fruto (los cuales están especificados para cada especie en el cuadro V) y comparar estos resultados con los encontrados en otros países que consideran a esta relación como indice para la cosecha en la industrialización de los cítricos. Se utilizaron tras arboles de naranja dulce criolla, identificados en este trabajo como : No. 28, 29 y 30; dos variedades de grape fruit: DUNCAN y FOSTER, dos variedades de mandarina; variedad carazo, cascara roja y variedad criolla, cascara amarilla y dos variedades de limón agrio: Criollo y Bear's Seedless. Los valores obtenidos de la relación sólidos totales solubles sobre acidez total son: naranja dulce del 9.2:1 a 9.3:1 (cuadro No. I), para la grape fruit FOSTER fue de 3.78:1, para la DUNCAN 6.6:1 (cuadro No. II); para la mandarina se obtuvo una relación de 21.1:1 a 18.6:1 (cuadro No. III) y para el limón ácido un porcentaje de ácido cítrico de 6.81 a 6.89 (cuadro No. IV). Estos resultados coinciden con los encontrados por Chandler (2) haciendo mención que en la naranja dulce, la relación sólidos/ácidos de este ensayo es mayor que la relación 8:1 encontrada en los Estados Unidos, debido quizás al mayor periodo cálido en que se desarrollan nuestros cítricos. Con respecto a la mandarina, la relación mencionada es mayor en la variedad Carazo, de color de cascara mas intenso. Este resultado coincide con el encontrado por Chandler (2). Esta relación no se encontró con la grape fruit. Con respecto al limon agrio, el porcentaje de ácido cítrico fue menor que el promedio de los resultados de FELIU (4), pudiendo deberse la diferencia al factor climático, tal como se explico con la naranja dulce. Finalmente se recomienda que estos resultados sean acompañados con un estudio de la palatabilidad de las frutas.

Interim report on the ground-water resources of Manatee County, Florida

A large part of western Manatee County is devoted to the growing of winter vegetables and citrus fruits. As in most of peninsular Florida, rainfall in the county during the growing season is not sufficient for crop production and large quantites of artesian water are used for irrigation. The large withdrawals of artesian water for irrigation result in a considerable decline of the artesian head in the western part of the county. This seasonal decline of the artesian head has become larger as the withdrawal of artesian water has increased. The lowering of the fresh-water head in some coastal areas in the State has resulted in an infiltration of sea water into the water-bearing formations. The presence of salty water in the artesian aquifer in parts of the coastal area of Manatee County indicates that sea water may also have entered the waterbearing formations in this area as a result of the decline of artesian pressure during the growing season. The purpose of the investigation is to make a detailed study of the geology and ground-water resources of the county, primarily to determine whether salt-water encroachment has occurred or is likely to occur in the coastal area. (PDF contains 38 pages.)

Rapid changes in the vegetation of a shallow pond in Epping Forest, related to recent droughts

Over much of Britain, 1995 and 1996 have been perceived as drought years. To evaluate the impact that local climatic conditions are having upon successional changes in higher vegetation (macrophytes), Speakmans Pond in Epping Forest was surveyed and mapped in 1996. The results are related to previous vegetation surveys carried out in 1989 and 1991. In 1989 the dominant marginal vegetation was floating sweet-grass Glyceria fluitans, which also covered a major part of the main body of the pond. Other abundant species included soft rush Juncus effusus, reed mace Typha latifolia and yellow flag Iris pseudocorus. A small (central) area of open water contained bladderwort Utricularia vulgaris and white water-lily Nymphaea alba. A similar plant coverage was found in 1991, with a dominance of floating sweet-grass along the shallow eastern edge. A marked change in the pond was found during the 1996 survey of vegetation in July, when the pool was dry. The major plant cover now consisted of creeping bent Agrostis stolonifera, with isolated clumps of Yorkshire fog Holcus lanatus around the edges; both are terrestrial grasses found on land surrounding the pond. Rushes (Juncus) had increased their distribution round the margins of the pond, and the patch of yellow flag noted in 1989 and 1991 was not found in 1996. The deeper trenches were also dry, but a small patch of white water-lily remained adjacent to one of the trenches.

How resources control aggression in Drosophila

How animals use sensory information to weigh the risks vs. benefits of behavioral decisions remains poorly understood. Inter-male aggression is triggered when animals perceive both the presence of an appetitive resource, such as food or females, and of competing conspecific males. How such signals are detected and integrated to control the decision to fight is not clear. Here we use the vinegar fly, Drosophila melanogaster, to investigate the manner in which food and females promotes aggression.

In the first chapter, we explore how food controls aggression. As in many other species, food promotes aggression in flies, but it is not clear whether food increases aggression per se, or whether aggression is a secondary consequence of increased social interactions caused by aggregation of flies on food. Furthermore, nothing is known about how animals evaluate the quality and quantity of food in the context of competition. We show that food promotes aggression independently of any effect to increase the frequency of contact between males. Food increases aggression but not courtship between males, suggesting that the effect of food on aggression is specific. Next, we show that flies tune the level of aggression according to absolute amount of food rather than other parameters, such as area or concentration of food. Sucrose, a sugar molecule present in many fruits, is sufficient to promote aggression, and detection of sugar via gustatory receptor neurons is necessary for food-promoted aggression. Furthermore, we show that while food is necessary for aggression, too much food decreases aggression. Finally, we show that flies exhibit strategies consistent with a territorial strategy. These data suggest that flies use sweet-sensing gustatory information to guide their decision to fight over a limited quantity of a food resource.

Following up on the findings of the first chapter, we asked how the presence of a conspecific female resource promotes male-male aggression. In the absence of food, group-housed male flies, who normally do not fight even in the presence of food, fight in the presence of females. Unlike food, the presence of females strongly influences proximity between flies. Nevertheless, as group-housed flies do not fight even when they are in small chambers, it is unlikely that the presence of female indirectly increases aggression by first increasing proximity. Unlike food, the presence of females also leads to large increases in locomotion and in male-female courtship behaviors, suggesting that females may influence aggression as well as general arousal. Female cuticular hydrocarbons are required for this effect, as females that do not produce CH pheromones are unable to promote male-male aggression. In particular, 7,11-HD––a female-specific cuticular hydrocarbon pheromone critical for male-female courtship––is sufficient to mediate this effect when it is perfumed onto pheromone-deficient females or males. Recent studies showed that ppk23⁺ GRNs label two population of GRNs, one of which detects male cuticular hydrocarbons and another labeled by ppk23 and ppk25, which detects female cuticular hydrocarbons. I show that in particular, both of these GRNs control aggression, presumably via detection of female or male pheromones. To further investigate the ways in which these two classes of GRNs control aggression, I developed new genetic tools to independently test the male- and female-sensing GRNs. I show that ppk25-LexA and ppk25-GAL80 faithfully recapitulate the expression pattern of ppk25-GAL4 and label a subset of ppk23⁺ GRNs. These tools can be used in future studies to dissect the respective functions of male-sensing and female-sensing GRNs in male social behaviors.

Finally, in the last chapter, I discuss quantitative approaches to describe how varying quantities of food and females could control the level of aggression. Flies show an inverse-U shaped aggressive response to varying quantities of food and a flat aggressive response to varying quantities of females. I show how two simple game theoretic models, “prisoner’s dilemma” and “coordination game” could be used to describe the level of aggression we observe. These results suggest that flies may use strategic decision-making, using simple comparisons of costs and benefits.

In conclusion, male-male aggression in Drosophila is controlled by simple gustatory cues from food and females, which are detected by gustatory receptor neurons. Different quantities of resource cues lead to different levels of aggression, and flies show putative territorial behavior, suggesting that fly aggression is a highly strategic adaptive behavior. How these resource cues are integrated with male pheromone cues and give rise to this complex behavior is an interesting subject, which should keep researchers busy in the coming years.

A colorimetric determination of hydrocyanic acid

The object of this investigation is to devise a rapid, fairly accurate, colorimetric analysis for HCN to be used in field work for determining instantaneous concentrations of the gas under fumigating canvas. A large amount of money is expended yearly by the citrus industry of this state in attempting to control and to eradicate the scale pests. Although fumigation with HCN has been practiced tor many years, the progress made has been anything but satisfactory. The greater portion of the work has always been carried on by contractors, who in a large number of cases have been very unscrupulous. The materials and labor are very expensive and the growers have been satisfied to adhere to beaten paths and hope for the best results on scale kill with the least attendant foliage injury. One familiar with fumigating, either from the grower's or the operator's viewpoint, knows that very widely varying results are obtained, even under what are apparently identical condition. Even after discounting for the dishonesty of some operators and the prejudices of the grower, there is still a large variance between desired or expected results and those actually obtained.

Cultura de tecidos e análise do potencial antioxidante de Passiflora alata Curtis

Passiflora alata Curtis, comumente conhecida como maracujá-doce, é uma das espécies do gênero Passiflora cultivadas comercialmente, sendo consumida in natura devido ao seu gosto adocicado. Ela também é utilizada em todo o mundo como ornamental e na medicina popular. O objetivo deste trabalho foi o estabelecimento de diferentes estratégias para a cultura in vitro de P. alata e a análise da produção de substâncias antioxidantes nos materiais obtidos in vitro, em comparação com as plantas in vivo. Diferentes tratamentos visando à quebra da dormência das sementes foram avaliados para a germinação in vitro ou in vivo, além da incubação das sementes sob tipos distintos de luz. Para o estabelecimento das culturas primárias, apices caulinares e segmentos nodais das plântulas derivadas da germinação in vitro foram cultivados em meio MSM . A taxa de alongamento dos brotos e o número de nós por brotos das culturas primárias foram aumentados pela adição de água de coco ao meio. Plantas derivadas dessas culturas foram utilizadas como fontes de explantes nodais, internodais e foliares. O potencial morfogênico de sementes sem tegumento foi também avaliado. Calos friáveis foram induzidos a partir de segmentos nodais e foliares na presença de PIC, e aqueles obtidos a partir de folhas em meio suplementado com PIC a 28,9 μM foram selecionados para o estabelecimento de culturas de células em suspensão. Após o desenvolvimento de diferentes estratégias in vitro para P. alata, folhas de plantas in vivo foram utilizadas para a avaliação de parâmetros que afetam a extração de substâncias antioxidante. O potencial antioxidante foi determinado pelo ensaio DPPH e o conteúdo de fenóis totais foi determinado utilizando o método Folin-Ciocalteau. Após o desenvolvimento do protocolo de extração, a atividade antioxidante dos diferentes materiais in vitro foi também avaliada. A eficiência antirradicalar variou entre os sistemas de cultura estudados, sendo diretamente proporcional ao conteúdo de fenóis totais dos extratos. Esses resultados indicam que as estratégias para cultura in vitro de P. alata desenvolvidas neste trabalho representam alternativas para a multiplicação de plantas e produção de substâncias fenólicas com ação antioxidante.

A Construção da História em Serena, de Ian McEwan

Esta dissertação constitui uma reflexão sobre a situação social, política e cultural da Inglaterra na década de 70, especialmente nos anos de 1972 e 1973, como descrita no romance Serena, de Ian McEwan. O objetivo principal desta pesquisa é apresentar como o momento histórico supracitado é narrado no discurso ficcional, a partir da perspectiva da personagem Serena, em comparação com o discurso histórico, de acordo com fontes selecionadas, que narram a história da Inglaterra no contexto da Guerra Fria, dos ataques terroristas do IRA e do funcionamento do Serviço Secreto doméstico inglês, chamado MI5.

甘薯体细胞胚胎发生和原生质体培养及转化的研究

甘薯[Ipomoea batatm L.]胚性愈伤组织的诱导、生长及分化，受遗传背景、外植体来源、激素配比等多种因素的影响。以徐薯l8叶片为外植体，在Ms附加2mg／l2.4-D的培养基上，诱导、筛选出三种类型的愈伤组织（I型、Ⅱ型、IV型），虽然其来源和培养条件相同，但在外部形态、内部结构、分化途径、分化能力、同工酶酶谱、可溶性蛋白质组成以及DNA分子结构方面，都有显著差异。I型愈伤组织在Ms无激素培养基上分化出体细胞胚，其过氧化物酶及脂酶同工酶与Ⅱ型愈伤组织和lV型愈伤组织相比，酶带的数目、深浅不同，而与胚状体相类似，可以作为不回类型愈伤组织及其分化途径的生理指标。SDS-PAGE电泳分析表明：三种愈伤组织的总蛋白组成也有显著差异。I型愈伤组织与Ⅱ型愈伤组织相比，无论是酶带的数目，还是酶带的扫描高度，都占绝对优势，表明其具有相对较高的蛋白质合成代谢水平。RAPD分析表明，三种类型愈伤组织的遗传基础具有一定差异。在所用的10个随机引物中，引物G3(GAGCCCTCCA)扩增出了显著的多态性，在I型愈伤组织与胚状体中发现两个特异片段，有可能与体细胞胚胎发生过程特异相关。 以I型愈伤组织进行悬浮培养，建立了具有再生能力的胚性悬浮细胞系，并对其鲜重、干重、PCV体积、PH值等生长特性进行了测定，研究了在悬浮培养条件下，2.4-D浓度对体细胞胚胎发生的影响。在MS附加Img/12.4-D的培养基中，细胞呈园球型，细胞质浓厚，细胞核显著，分裂旺盛，转入不含2.4-D的分化培养基中，即可得到大量的游离胚状体：浓度过高（4-8mg／l）或过低（0.5mg/l），都不利于细胞的生长和分化。胞外同工酶研究发现：胞外过氧化物酶水平，随着2.4-D浓度升高、培养时间的延长、细胞生长速度的减缓而降低，随着体胚分化的开始而升高。这种变化趋势表明：2.4-D浓度、过氧化物酶及细胞的生长、分化之间，存在密切联系。蛋白质分析发现，2.4-D的浓度与细胞内可溶性蛋白质组成及含量也密切相关：在无激素培养基中的培养物，其蛋白质电泳图谱与在含有不同浓度2.4-D的培养基中的培养物相比，无论是谱带的数目还是谱带的峰值，都有深刻差异，表明在体胚分化和发育过程中，细胞内进行着旺盛的蛋白质合成代谢。 以胚性悬浮细胞为材料，进行原生质体培养。通过对游离条件的比较研究，发现采用PH值为6.0的E3酶液酶解2小时，可以得到大量具有旺盛活力的原生质体。采用液体浅层一固体平板双层培养方式进行培养，原生质体分裂旺盛，20天后形成肉眼可见的微型愈伤组织，在附加0.5mg/1 2.4-D的继代培养基中，出现根的分化，转入附加0.5mg/12.4-D和Img/16-BA的分化培养基中，有少量不定芽发生，并形成小苗，但无胚状体产生。 以来源于胚性悬浮细胞的原生质体为受体，通过与农杆菌共培养，将苏云金杆菌毒蛋白(Bt)基因导入甘薯细胞，经卡那霉素筛选，得到抗性愈伤组织，并有根的分化。经过PCR检测，证明了Bt基因在甘薯细胞染色体组中的整合。

甜樱桃果实采后生理、耐藏性及褐变机理的研究

本文以我国目前主要栽培的甜樱桃早、中、晚熟品种“红灯”、“佐藤锦”、“那翁”、“拉宾斯”和“砂蜜豆”等为试材，系统地研究了甜樱桃果实在不同O2和CO2浓度气调贮藏条件下的生理特性、风味品质和贮藏性，提出了适合于甜樱桃果实生理特性的气调指标和贮藏时间;分析了甜樱桃在不同贮藏条件下果实中乙烯、多酚氧化酶（PPO）、过氧化物酶（POD）、超氧化物歧化酶（SOD） 和丙二醛（MDA）的变化情况，揭示了它们与甜樱桃果实衰老、褐变及耐贮性的关系;分析了甜樱桃果实在不同贮藏条件下乙醇、色泽、硬度、可溶性固形物、酸和维生素C等物质成分的含量变化及与果实风味品质的关系;同时，还分析了不同甜樱桃品种果皮的结构特征及与耐藏性的关系，以及不同成熟度的甜樱桃果实在气调贮藏中的生理反应和品质变化。为形成甜樱桃果实采后商业化贮藏的系列配套技术提供了理论依据。 本文的主要研究内容包括：不同CO2浓度的气调贮藏对甜樱桃果实采后生理及品质的影响;高O2或高CO2浓度处理对甜樱桃果实褐变、衰老及风味品质的影响;动态气调贮藏和非动态气调贮藏对甜樱桃生理特性、风味品质和耐藏性的影响;不同成熟度及不同品种对甜樱桃耐藏性的影响;不同药剂配合包装处理对甜樱桃果实耐藏性及品质的影响。试验结果表明： 1. 甜樱桃果实适合于较高CO2浓度的CA贮藏：与自发性气调贮藏（MAP）相比，气调贮藏（CA）更能明显地延缓果实衰老，减少腐烂和褐变，保持风味品质和延长1.5~2倍的贮藏时间。其中以较高CO2浓度的（5% O2 + 10% CO2）气调贮藏效果最好，早熟品种“红灯”的贮藏期为60天，而中、晚熟品种“最上锦”、“佐藤锦”、“那翁”、“拉宾斯”和“砂蜜豆” 的贮藏期可达80-100天。果实在25℃下的货架存放时间为2-3天，在低温条件下（3-5℃）的货架时间是6天以上。 2. 高O2浓度气调对甜樱桃果实易造成伤害：用高O2浓度（70% O2 + 0% CO2）气调贮藏甜樱桃，在短期内可以抑制果实腐烂、减少果肉中挥发性代谢产物乙醇含量，但果实中的丙二醛含量上升迅速，褐变加剧，造成高氧伤害。 3. 动态气调贮藏更有利于延长甜樱桃果实的贮藏期：与非动态气调贮藏相比，贮藏前期用较高浓度CO2（20%）处理甜樱桃果实可有效控制维生素C含量的降低，丙二醛含量上升的速率较慢，可明显减少贮藏后期果实褐变和腐烂，保持风味品质。 4. 成熟度较高的甜樱桃果实适合于气调贮藏：成熟度较高（深红色时采收）的甜樱桃果实比成熟度低（鲜红色时采收）的果实耐藏性好。 5. 果实的耐藏性与果皮结构有关：耐藏性强的品种表皮细胞较小，排列紧密，细胞壁和外层蜡脂均较厚，果肉内部的维管束螺纹排列致密，果肉的薄壁细胞也排列得比较致密。 6. MAP与生物药剂配合有利于延长甜樱桃的贮藏期：生物1号药剂与加有TBZ的低密度塑料薄膜袋包装配合使用对控制甜樱桃果实腐烂、抑制果肉乙烯和乙醇生成、保持品质特性和延长贮藏时间都很有效。 7. 膜质过氧化是造成甜樱桃果实褐变的主要因素：甜樱桃果实褐变与PPO活性关系不大，但与膜质过氧化作用的产物—丙二醛（MAD）含量变化显著相关。

棉花抗黄萎病基因工程研究和防卫反应相关基因的克隆

系统获得性抗性(Systematic acquired resistance, SAR)是植物抵御病原菌侵染的最有效手段，利用基因工程技术导入SAR信号发生过程中的关键基因后，植物的SAR基因表达量提高并且对病原菌侵染的反应速度加快，因此植物的抗病性得以增强，与传统的抗病基因工程技术相比它对病原菌没有专一性，许多学者称之为广谱抗病基因工程，该领域已成为目前抗病基因工程研究的热点和前沿。 NDR1和NPR1基因在植物的SAR发生中起着重要的作用，前者功能定位在ROS(reactive oxygen species)的激活和随后的水杨酸(SA)诱导合成之间，突变株病原菌诱导后SA合成能力降低，SAR发生减弱，目前还没有对该基因进行过量表达分析的报道;后者功能定位在SAR信号转导级联反应之中的SA积累和随后的SAR基因表达之间。该突变株在病原菌侵染时不产生病程相关蛋白(PRs)，表现为感病，而对照抗病;过量表达该基因的转基因拟南芥对多种病原菌的侵染产生抗性，PR1等PRs蛋白的表达量也提高，异源表达该基因的水稻对白叶枯病的抗性也提高。本研究利用RT-PCR方法从拟南芥中克隆了这两种基因，序列分析表明拟南芥Wassilewskija生态型的NDR1基因与Columbia生态型相比，共有7处碱基不同，引起编码氨基酸变化4处，而NPR1基因与报道的Wassilewskija生态型来源的NPR1基因完全相同。 我们构建了35S启动子驱动的NDR1和NPR1基因的植物组成型高效表达载体，利用农杆菌介导法转化烟草，PCR和Southern鉴定外源基因已经整合到植物基因组中。抗病性分析显示过量表达NDR1和NPR1基因的烟草对晚疫病和赤星病的抗性都有明显提高，说明这两个基因的在其它植物中异源表达后，都能提高植物对多种病原菌的抗性。 本论文提出了利用这两个基因来培育抗黄萎病棉花的设想，一方面为解决这个“世纪性”难题积累新的资料，另一方面也为其它作物的抗病基因工程提供新的经验。利用35S启动子驱动的NDR1和NPR1基因的植物组成型表达载体分别对陆地棉品种石远321进行花粉管通道法转化。同时，还探讨了这两个基因在棉花中的共转化实验，希望它们的“协同增效”能进一步提高棉花的抗病性。对其中2001年夏天在南京注射所获得的5,000粒种子在三亚进行100 g/ml卡那霉素筛选，初步鉴定分别获得转NDR1和NPR1基因株系26和24棵，PCR进一步鉴定其中分别有12和7棵为转基因阳性，转基因频率分别为0.50％和0.27％，目前利用营养钵蘸根法对其二代进行抗枯、黄萎病鉴定，结果显示有转基因植株对枯、黄萎病的抗性都明显增强，进一步的鉴定正在进行中。2002年初海南注射分别获得转NDR1和NPR1基因以及共转化种子22,000、10,500和12,500粒种子，2002年夏在中国农科院植保所黄萎菌病圃筛选抗黄萎病单株，并利用100 g/ml卡那霉素初步筛选出了一批抗性植株，每种转基因株系随机挑选5株进行PCR鉴定，结果显示为阳性。进一步的抗黄萎病鉴定和筛选以及分子分析正在进行中。 同时，本文还探讨了病原菌诱导型启动子在广谱抗病基因工程应用的可能性。根据烟草的Pr1-a启动子已知序列设计引物，PCR扩增启动子序列后,构建病原菌诱导型NPR1基因植物表达载体，并对棉花进行转化，获得种子11,500粒，利用同上的筛选方法，获得了一致的结果，目前抗黄萎病鉴定、分子检测以及生物学分析正在进行中。 最后，鉴于抗生素标记在转基因植物的应用引起了许多“安全性”争论的事实，还构建了无筛选标记的表达载体对抗虫棉进行转化，这样在生产上可以直接获得抗虫棉抗黄萎病棉花新材料，也为其它作物抗病基因工程积累经验。 本研究还提出了一种较为有效的提取高质量棉花总RNA的方法，与原来一些棉花RNA纯化方法相比，该方法所用都为常规试剂，易于重复，质量高。并且利用获得的总RNA构建了黄萎菌激发子诱导的cDNA文库，滴度测定为1╳107pfμ/μg，插入片段大小在5 00~2 000 bp范围内。 鉴于NPR1基因研究的重要性，本研究还利用简并引物PCR技术从海岛棉和陆地棉的基因组中都分离到了NPR1基因的同源片段，大小都为208 bp，与拟南芥NPR1基因的相应部分的同源性分别为66％和65％，它们之间的同源性为87％，目前该基因的全长正在分离鉴定中。 多聚半乳糖醛酸酶抑制蛋白(PGIPs)在植物的防御反应中起着重要的作用，通过分析已知20余种pgip基因序列的保守区，设计简并引物，PCR扩增海岛棉（Gossypium barbadense）7124 cDNA文库，得到一条长561 bp的片段，序列测定后分析确认为pgip基因的一部分。根据此序列和棉花病原菌诱导的cDNA文库载体中已知部分设计RACE引物，扩增后，5’和3’RACE分别得到666bp和906 bp的片段。序列分析表明它具有完整的编码框，产物为330 aa的蛋白质。序列分析该蛋白具有10个串联的LRR(leucine-rich repeat)区，与柑桔(Citrus)和枳(Poncirus)的pgip基因的同源性分别为69.2%和68.7%。进一步PCR扩增得到该基因的全长阅读框，并且获得了相应的基因组片段，序列分析发现该基因没有内含子。这是从棉属植物中克隆的第一个pgip基因。

大功率微波与臭氧等复合因子对大棚蔬菜根结线虫及土壤的作

虫（nematodes）是世界农业生产的一大障碍，每年给世界农业生产造成约1000亿美元的巨额损失，是世界各国一直关注的重点病害之一。线虫可以危害各种大田作物和各种温室植物，几乎所有的栽培植物都有线虫危害发生。随着人们环保意识的增强以及健康意识的觉醒，传统防治根结线虫的药物如溴甲烷等因对环境的破坏作用，或者由于高毒、使用不便、抗药性等原因而迅速退出历史舞台，研究环保高效防治线虫成为世界各国竞相发展的技术之一。本研究尝试通过物理方式防治蔬菜根结线虫，并调查了对土壤质量的影响，主要内容包括： 1. 设计制造了功率20 Kw，频率为915 MHz的大功率土壤微波处理机，并通过了田间试验; 2. 设计制造了每小时120 g臭氧产量的便携式臭氧土壤处理机，并通过了田间试验; 3. 采用4因素3水平正交法L9(34)在山东寿光蔬菜大棚进行了蔬菜根结线虫的防治试验，综合考察微波、臭氧、微波吸收剂、EM菌等不同处理水平对根结线虫和对土壤质量的影响，结果表明： 1) 大棚土壤经微波60 s照射能够显著降低甜瓜根结线虫数和根结指数（p < 0.05），盆栽试验只需处理10 s即可显著防治甜瓜根结线虫（p < 0.01）;以120 g/h臭氧浓度处理5 s也能显著降低甜瓜根结线虫的数量（p < 0.05）;田间施入5 g/m2的EM菌，能够显著降低甜瓜根结线虫的数量（p < 0.05）;盆栽试验中微波吸收剂具有降低根结线虫数量的趋势，但是不同处理水平之间差异不显著，田间试验则具有增加根结线虫的趋势，不同处理水平之间也没有显著差异。 2) 微波不同处理水平之间对甜瓜单瓜重的影响没有显著差异;以120 g/h臭氧浓度处理甜瓜能够显著增加甜瓜单瓜重（p < 0.05）;200 g/m2微波吸收剂能够显著提高甜瓜单瓜重（p < 0.05）;以5 g/m2EM菌处理土壤则显著降低甜瓜单瓜重（p < 0.05）。 3) 微波、臭氧、微波吸收剂不同处理水平对甜瓜糖度的影响没有显著差异;以3 g/m2的EM菌处理甜瓜能够显著降低甜瓜的糖度。 4) 不同因素对甜瓜硬度的影响没有显著差异。 5) 微波处理60 s能够显著提高甜瓜的果型指数（p < 0.1），臭氧、微波吸收剂、EM菌等因素不同处理水平之间对果型指数影响差异不显著。 6) EM菌5 g/m2处理浓度能够显著降低甜瓜株高（p < 0.1），微波、臭氧、微波吸收剂不同处理水平之间对株高的影响没有显著差异。 7) EM菌5 g/m2处理浓度能够显著降低甜瓜根径（p < 0.1），微波、臭氧、微波吸收剂不同处理水平之间对根径的影响没有显著差异。 8) 微波处理60 s能够显著降低表层土壤（0-5 cm）的有机质含量（p < 0.01）;以120 g/h的臭氧浓度处理，则可以显著提高10-15 cm土壤的有机质含量（p < 0.05），其它处理因素的不同水平对有机质的影响差异不显著。 9) 微波不同处理水平对土壤全氮的影响没有显著差异（p < 0.05）;120 g/h臭氧处理浓度能够显著降低表层土壤（0-5 cm）的土壤全氮量（p < 0.1）;100 g/m2微波吸收剂处理水平能够显著提高10-15 cm土壤全氮量（p < 0.05）;EM菌3 g/m2的处理水平则显著降低5-10 cm土壤全氮量（p < 0.05）。 10) 微波、微波吸收剂、EM菌不同处理水平之间对土壤全磷没有显著影响;240 g/h臭氧浓度处理5 s能够显著降低10-15 cm土层的土壤全磷含量(p < 0.05)。 11) 微波、微波吸收剂对土壤全磷没有显著影响;臭氧处理具有降低土壤有效磷的趋势，EM菌处理则有提高土壤有效磷的趋势，但是 不同处理水平之间差异不显著（p > 0.05）。 12) 微波处理对土壤EC值没有显著影响;臭氧、微波吸收剂、EM菌处理具有降低土壤EC值的趋势，但是不同处理水平之间差异不显著（p > 0.05）。 13) 微波处理30 s能够显著降低表层土壤的pH值（p < 0.05），臭氧、微波吸收剂、EM菌处理具有提高土壤pH值的趋势，但是不同处理水平之间差异不显著。