Detection of antigenic proteins expressed by lymphocystis virus as vaccine candidates in olive flounder, Paralichthys olivaceus (Temminck & Schlegel)

Although the major capsid proteins (MCPs) of lymphocystis disease virus (LCDV) have been characterized, little is known about the host-derived immune response to MCPs and other LCDV antigenic proteins. To identify antigenic proteins of LCDV that could be used as vaccine candidates in olive flounder, Paralichthys olivaceus, we analysed the viral proteins responsible for its virulence by applying immuno-proteomics. LCDV proteins were separated by one-dimensional gel electrophoresis, transferred to polyvinylidene difluoride membrane, and probed with homogeneous P. olivaceus antisera elicited by LCDV natural infection and vaccination with formalin-killed LCDV. Four immune-reactive proteins were obtained at 68-, 51-, 41- and 21 kDa using antisera collected from natural infection while two proteins at 51- and 21 kDa exhibited response to antisera from vaccinated fish, indicating that the latter two proteins have vaccine potential. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nanoelectrospray MS/MS, the 51 and 21 kDa proteins were identified as MCP and an unknown protein, respectively.     

Effects of modulatory agents on neurally-mediated responses of trout intestinal smooth musclein vitro

Mediators and mechanisms responsible for the inhibitory modulation of trout intestinal smooth muscle were examined using a series of putative mediators and substances known to modulate neurotransmission in mammalian systems. Frequency response relationships to transmural stimulation and concentration response relationships to 5-hydroxytryptamine, carbachol, and substance P were established on paired segments of rainbow trout intestinein vitro in the presence and absence of putative modulatory agents. Modulation of neurally-mediated contractions of trout intestine was achieved with dibutyryl cyclic AMP and forskolin, agents that increase intracellular levels of cyclic AMP. The effect appears to be at the level of the smooth muscle, since the adenylate cyclase activator, forskolin, inhibited muscarinic and serotoninergic contractions as well as transmurally stimulated contractions. Substance P-induced contractions were unaffected by forskolin. The endogenous agonists/neurotransmitters which would increase cyclic AMP levels in rainbow trout intestinal smooth muscle are as yet unknown. The effects do not appear to be modulated by vasoactive intestinal peptide (VIP), calcitonin, calcitonin gene-related peptide (CGRP), or agents that activate -adrenoceptors. Prostaglandin E₂ (PGE₂) and ₂-adrenergenic agonists are possible agents which will decrease contractility of the smooth muscle. They were only active in the proximal intestine and on transmurally stimulated contractions. The effects of both PGE₂ and ₂-agonists appear to be prejunctional, decreasing release of contractile neurotransmitters in the enteric nervous system.     

Effects of steroid hormones on immunoglobulin M (IgM) in rainbow trout, Oncorhynchus mykiss

The immunosuppressive effects of steroid hormones were evaluated as the response against implanted steroid hormones, cortisol (F), testosterone (T), estradiol-17 (E2), and 11- ketotestosterone (11-KT), in juvenile rainbow trout. In long term experiments (5 weeks), fish were given a single intraperitoneal implant of F or T. A clear suppressive effect of plasma IgM levels with F and T was not necessarily obtained, although mucus IgM levels were reduced corresponding to the elevated plasma steroid hormone levels. In short term experiments (1 week), intraperitoneal implantation of T, 11-KT and E2 suppressed plasma and mucus IgM levels, although the effects were not dose-dependent. When administered through diet, F and T caused a suppression of plasma IgM levels; F administration at both high and low dosages caused a significant decrease in plasma IgM levels, while only a high dose of T caused the suppression. These results suggest that sex steroid hormones, as well as F, have immunosuppressive functions in rainbow trout.     

Binding characteristics and regulation of the 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) receptor on testicular and sperm plasma membranes of spotted seatrout (Cynoscion nebulosus)

The binding characteristics of 17,20,21-trihydroxy-4-pregnen-3-one (20-S) to plasma membranes prepared from the testes and sperm of spotted seatrout (Cynoscion nebulosus) were investigated using a filtration method to retain the bound 20-S. A single class of high affinity (K_d = 17.9 nM), low capacity (B_max = 0.072 nM g^-1 testes) binding sites was identified by saturation and Scatchard analyses on testicular membranes of spermiating spotted seatrout. A corresponding receptor (K_d = 22.17 nM, B_max = 0.00261 nM ml^-1 milt) was also detected in spermatozoan membrane preparations. The rates of 20-S association and dissociation were rapid, both had T_halfs of less than 1 min. Competition studies indicated that the receptor was highly specific for 20-S. 17,20-dihydroxy-4-pregnen-3-one, which had the highest affinity of the other steroids tested, had a relative binding affinity (RBA) of 14.3%. Progesterone, 11-deoxycortisol and testosterone competed with an order of magnitude less affinity (RBA's of 7.4, 1.8 and 1.1%, respectively). Estradiol displayed low affinity for the receptor (RBA = 0.4%) and cortisol did not cause any displacement at 1000-fold excess concentration. Specific 20-S receptor binding was detected in plasma membranes from testes of both spermiating and non-spermiating seatrout and on spermatozoa. Prolonged incubation of testicular fragments from a spermiating fish with gonadotropin (15 IU ml^-1 human chorionic gonadotropin) or forskolin (10 µM) caused a 2–3 fold increase in membrane receptor binding. Previous studies have shown that gonadotropin-induced upregulation of the 20-S plasma membrane receptor in seatrout ovaries is required for the oocytes to become responsive to 20-S and undergo final maturation. The existence of a 20-S membrane receptor on sperm and its upregulation in the testes by gonadotropin raises the possibility that final maturation of spermatozoa in male seatrout may be regulated by a similar mechanism.     

中国烟草种质资源网络信息系统的开发

为促进中国烟草种质资源的共享与应用,研究开发了基于B/S（browser/server）架构和ASP.NET技术的中国烟草种质资源网络信息系统。数据库采用了Microsoft SQL Server,服务器端程序集应用C#语言开发,浏览器端基于XHTML + JavaScript。系统在特性数据库和共性数据库两个底层数据库的基础上,实现了种质数据查询、核心种质查询、种质图像查询、共性数据查询和烟草种质共享等5种功能。该系统突破了长期以来种质资源信息、数据、实物集中于一处,难以共享的局面,促进了种质资源的广泛应用和深入发展。     

玉米芯和桉木的低温热解特性

为实现生物质资源的分级综合利用,该文采用热重分析仪和裂解气质联用仪进行了对玉米芯和桉木低温热解特性的研究。试验结果表明不同生物质原料低温快速热解产物有明显差异,玉米芯的低温快速热解产物主要有乙酸、2,3-二氢-苯并呋喃和2-甲氧基-4-乙烯基苯酚,而桉木的产物主要是乙酸、糠醛和5,6-二氢-4-羟基-吡喃-2-酮。生物质低温快速热解产物种类较少,分布较为集中,玉米芯和桉木的酸类、呋喃类,桉木的吡喃类热解产物相对含量随温度上升而降低。生物质低温热解能有效分解其半纤维素,这为降低中温热解油的酸性和水分提供了理论指导。     

中国“产业沼气”的开发及其应用前景

“产业沼气”是指规模化地生产并纯化沼气,使之能像天然气和液化气那样,由管道或气罐（瓶）输送和装运,方便地应用;有别于通常所说的农户沼气。中国天然气资源很紧缺,而能替代天然气的沼气的巨大资源潜力尚未被广泛认识和认真地开发。该文分析对比了欧盟国家开发产业沼气的成功实践,提出要使中国有机废物/液的沼气处理摆脱只讲环保公益、缺乏经济效益的被动局面,开发出可观的洁净商品沼气,实现环境效益和经济效益“双赢”。该文还分析了中国产业沼气的5种主要资源和实现产业化必须克服的技术、政策瓶颈。结论指出,中期内中国产业沼气具有相当于年产900亿m3天然气的开发潜力,是很有前景的生物质能源。     

不同植物LDOX/ANS基因的生物信息学分析

花青素是一类重要的植物次生代谢产物。本文采用生物信息学的方法对已在GenBank上登录的拟南芥、西洋梨、苹果、桃、葡萄、芥菜、菊花新品种和水稻等植物的无色花青素双加氧酶/花青素合成酶(LDOX/ANS)基因的核酸及氨基酸序列、组成成分、导肽、信号肽、跨膜结构域、疏水性/亲水性、蛋白质二级结构、三级结构及功能域等进行预测和推断。结果表明,ORF除了桃和菊花新品种为500bp左右之外,其它几种植物都在1070bp左右,分子量均为4kD左右,理论等电点均低于7,说明LDOX/ANS呈酸性。Glu、Leu和Val是这些植物共有的主要氨基酸。核苷酸同源性比对结果显示,拟南芥LDOX/ANS与其它植物LDOX/ANS的同源性较高;8个物种的LDOX/ANS基因被分为两个大类,单子叶植物水稻LDOX/ANS基因被单独分为一类,其它的均聚成一大类。研究还发现这些植物的LDOX/ANSN端不存在导肽和信号肽,无跨膜结构域,肽链表现为亲水性。蛋白质二级结构中最主要的结构元件是α-螺旋和无规则卷曲,包含一个20G-FeⅡOxy功能结构域。通过此次研究,希望为今后深入研究该类酶的功能和结构特征提供依据。     

不同硝铵比对霞多丽葡萄幼苗生长和氮素营养的影响

采用沙培试验,设置了等氮条件下5种不同硝铵比营养液处理,探讨了氮形态对霞多丽葡萄幼苗生长和氮素营养的影响。结果表明,霞多丽幼苗的生长量以硝铵比70/30时最大,全铵营养时最低;植株根系氮的平均吸收量在硝铵比为70/30时为最大值,达到70.89%,全铵营养时为最低值;叶片中的氮浓度和总氮量以全硝营养最大,全铵营养最低,后者仅是全硝营养的61.8%和19.46%。霞多丽幼苗根系内NH4+-N浓度与营养液中NH4+-N的浓度成极显著正相关(r=0.9805),且当铵态氮比例超过30%时根系中NH4+浓度显著增加。叶片中硝酸还原酶的活性以硝铵比70/30最高,当铵态氮比例大于30%,再继续增加铵态氮的比例时,显著抑制了叶片中硝酸还原酶的活性;而对根的硝酸还原酶活性无明显影响。试验证明,硝铵比70/30是霞多丽较适宜的氮素形态配比,较高比例的铵态氮增加了植株器官中NH4+的浓度,抑制了霞多丽幼苗的生长及对氮素的吸收积累。     

Long-term organic phosphorus mineralization in Spodosols under forests and its relation to carbon and nitrogen mineralization

In forest soils where a large fraction of total phosphorus (P) is in organic forms, soil micro-organisms play a major role in the P cycle and plant availability since they mediate organic P transformations. However, the correct assessment of organic P mineralization is usually a challenging task because mineralized P is rapidly sorbed and most mineralization fluxes are very weak. The objectives of the present work were to quantify in five forest Spodosols at soil depths of 0-15 cm net mineralization of total organic P and the resulting increase in plant available inorganic P and to verify whether net or gross P mineralization could be estimated using the C or N mineralization rates. Net mineralization of total organic P was derived from the net changes in microbial P and gross mineralization of P in dead soil organic matter. We studied very low P-sorbing soils enabling us to use lower extractants to assess the change in total inorganic P as a result of gross mineralization of P in dead soil organic matter. In addition, to enable detection of gross mineralization of P in dead soil organic matter, a long-term incubation (517 days) experiment was carried out. At the beginning of the experiment, total P contents of the soils were very low (19-51 μg g⁻¹) and were essentially present as organic P (17-44 μg g⁻¹, 85-91%) or microbial P (6-14 μg g⁻¹; 24-39%). Conversely, the initial contents of inorganic P were low (2-7 μg g⁻¹; 9-15%). The net changes in the pool size of microbial P during the 517 days of incubation (4-8 μg g⁻¹) and the amounts of P resulting from gross mineralization of dead soil organic matter (0.001-0.018 μg g⁻¹ day⁻¹; 0.4-9.5 μg g⁻¹ for the entire incubation period) were considerable compared to the initial amounts of organic P and also when compared to the initial diffusive iP fraction (<0.3 μg g⁻¹). Diffusive iP corresponds to the phosphate ions that can be transferred from the solid constituents to the soil solution under a gradient of concentration. Net mineralization of organic P induced an important increase in iP in soil solution (0.6-10 μg g⁻¹; 600-5000% increase) and lower increases in diffusive iP fractions (0.3-5 μg g⁻¹; 300-2000% increase), soil solid constituents having an extremely low reactivity relative to iP. Therefore, soil micro-organisms and organic P transformations play a major role in the bioavailability of P in these forest soils. In our study, the dead soil organic matter was defined as a recalcitrant organic fraction. Probably because gross mineralization of P from this recalcitrant organic fraction was mainly driven by the micro-organisms’ needs for energy, the rates of gross mineralization of C, N and P in the recalcitrant organic fraction were similar. Indirect estimation of gross mineralization of P in dead soil organic matter using the gross C mineralization rate seems thus an alternative method for the studied soils. However, additional studies are needed to verify this alternative method in other soils. No relationships were found between microbial P release and microbial C and N releases.