昆虫病毒资源的开发及其应用进展

昆虫数量占地球上现存动物数量的65%,感染昆虫的病毒种类更是纷繁多样。与其他微生物相比,病毒具有基因组小、遗传操作简单等特点,是一类蕴藏巨大应用潜能的生物资源。在过去的十年里,随着新一代测序技术和生物信息学科的迅猛发展,科学界进入了一个快速鉴定昆虫新病毒的新时代。昆虫病毒多样性研究为揭示昆虫和病毒的协同进化机制提供新见解,同时为昆虫病毒的商业化应用带来新思路。本文根据昆虫病毒近期的研究进展,概述高通量测序技术鉴定昆虫病毒的原理和方法,并对昆虫病毒的资源属性及应用前景进行探讨,为有害昆虫的防治及虫媒病害的防控提供理论基础。     

病毒诱导的基因沉默及其在植物功能基因组研究中的应用

病毒诱导的基因沉默已成为研究植物功能基因组的重要工具. VIGS 体系因其方法简便、周期性短以及避免植物转化等诸多优点, 已在利用正向遗传学和反向遗传学寻找和鉴定基因功能方面发挥了日益重要的作用. 越来越多的植物病毒被改造成为VIGS 载体, 并已在植物发育、生物逆境、非生物逆境、细胞代谢、信号传导等基因功能研究方面得到了应用. 本文围绕VIGS的发展以及在植物功能基因鉴定中的应用及前景提出了展望.     

双链RNA技术在植物病毒研究中的应用

含ＲＮＡ基因组的植物病毒在复制时会产生双链ＲＮＡ（ｄｓＲＮＡ）。本文介绍了用ＣＦ－－１１纤维素粉提取ｄｓＲＮＡ的步骤,并列举了ｄｓＲＮＡ在植物病毒研究中的应用,其主要应用有（１）用于检测植物病毒;（２）用于病毒株系分化;（３）用于病毒卫星ＲＮＡ及亚基因组ＲＮＡ研究;（４）用于侵染性测定及病毒基因组功能研究等。     

植物病毒双链RNA提取、研究进展

植物病毒病在世界各地广泛分布,大约90%的植物病毒基因组为单链RNA,当病毒侵染植物后利用寄主成分进行复制时,首先产生与基因组RNA互补的链,配对成双链模板,再以互补链为模板转录出子代基因组RNA。而正常的植物中往往不产生,而且ds-RNA对酶具有一定的抗性。通过对植物组织中ds-RNA的分析,可用于植物病毒的检测和诊断等研究。因此,植株中病毒ds-RNA的分离和鉴定,对病毒的研究和基因组解析具有重要意义。     

植物病毒诱导的基因沉默在基因功能研究中的应用

传统的植物遗传转化方法周期长、工作量大、过程繁琐,不利于基因功能的快速高通量鉴定．近年来随着基因沉默机制研究的深入和不断发展,利用病毒诱导的基因沉默(Virus induced gene silencing,VIGS)进行植物功能基因组研究作为一种快速、高通量的反向遗传学工具已被广泛应用在烟草、马铃薯、番茄等植物中, 在大规模的植物基因组功能鉴定中展示了广阔的应用前景．综述了 VIGS 的作用机制、植物病毒栽体、转化方法以及在植物基因功能研究等方面的应用及前景．     

渗漏型UGA终止密码在植物病毒相关基因组中的作用

弱毒疫苗ToMV-K的复制酶基因在2670-2672核苷酸处发生UGA突变,研究表明该突变是导致病毒弱化的主要原因。通过ToMV-K复制酶的突变区与其它具有UGA渗漏终止密码的植物ssRNA病毒基因组通读结构区的分析和比较,发现ToMV-K和其它植物病毒的UGA渗漏与通读相关基因的共同特征：CGG基元,通读区的α-螺旋结构和一些疏水氨基酸残基使UGA的通读成为可能。这些渗漏与通读的特征可能才是ToMV\|K致弱的根本原因。可以根据这一模式,探讨对其它植物ssRNA的病毒如PVX、PVY、CMV等的基因组改造和致弱研究。     

重组病毒杀虫剂应用研究进展

应用分子生物学技术可以将昆虫特异性的毒素基因、某些酶基因等外源基因插入昆虫病毒基因组,或通过改造昆虫病毒基因组等方法构建重组病毒杀虫剂,提高杀虫效果。温室及田间释放实验证实,重组病毒杀虫剂可以显著提高现场防治效果。连续多代抗性筛选实验表明,宿主被诱导产生对重组病毒杀虫剂抗性的速度低于野生型病毒杀虫剂。采用在剂型中添加光增白剂等保护剂、在基因组中插入具有增效作用的基因、应用病毒增强蛋白等技术可以提高重组病毒杀虫效果。随着基因工程技术的发展和安全性研究的深入,以重组杆状病毒为主的重组昆虫病毒杀虫剂的应用研究正面临着突破。     

RNA沉默与植物病毒

植物中RNA沉默（RNAsilencing)亦称为转录后基因沉默（PTGS）或共抑制,是植物抵抗外来核酸（转座子、转基因或病毒）入侵,并保护自身基因组完整性的一种防御机制。RNA沉默是近十年来发现的植物界中普遍存在的现象,已成为植物分子生物学领域的一个新的研究方向。对RNA沉默特点和机制的研究表明,植物病毒与（转基因）植物内发生的RNA沉默有着密切的联系,作者从病毒对RNA沉默的诱导、抑制、防御等方面,简述了RNA沉默与病毒的关系。并对病毒载体所诱导的RNA沉默在植物发育和基因组功能分析等方面的应用价值进行了讨论。     

植物病毒载体系统研究进展

植物病毒基因组小,易于进行遗传操作而且感染过程简单,因而利用植物病毒载体表达外源基因在生物技术领域具有潜在的应用优势。本文主要介绍了抗原展示系统和多肽表达系统,并分别列举一些植物病毒作为表达载体的研究进展情况以及应用前景。     

植物病毒学的研究进展

1国内外植物病毒的研究进展 植物病毒是病毒学科的一个重要组成部分,从病毒学的发展历史来看,一些开创性的工作和基础理论研究成果最先是在植物病毒领域里取得的.这类病原物给很多重要经济作物造成巨大危害,全世界每年农作物的损失达200亿美元.20世纪初对植物病毒学的研究主要集中在植物病理学领域,随着社会的发展和科学技术的进步,植物病毒与生物化学、生物物理学、免疫学、医药学等相关学科相互影响、相互渗透,尤其是在生物化学、分子生物学、植物基因工程技术、信息科学等领域的飞速发展,以及新技术新方法的不断应用,病毒学发展焕然一新.我们的研究手段已经从细菌过滤器、电子显微镜、X射线衍射法、扫描投射电镜过渡到现代分子生物学技术的应用如分子克隆、重组、体外表达等;这些方法的应用使得研究学者们对植物病毒的研究从形态、结构、组成及病毒核酸和蛋白质大分子的超微结构的观察,深入到近年来的对植物病毒基因组核苷酸序列的分析和氨基酸组成及结构功能上,进一步揭示了一些植物病毒的本质特征,使人们对植物病毒有了更深刻地认识,无论是在理论上还是在应用上都有了很大发展.     

Viral Detection: Past,Present, and Future

Viruses are essentially composed of a nucleic acid (segmented or not, DNA, or RNA) and a protein coat. Despite their simplicity, these small pathogens are responsible for significant economic and humanitarian losses that have had dramatic consequences in the course of human history. Since their discovery, scientists have developed different strategies to efficiently detect viruses, using all possible viral features. Viruses shape, proteins, and nucleic acid are used in viral detection. In this review, the development of these techniques, especially for plant and mammalian viruses, their strengths and weaknesses as well as the latest cutting‐edge technologies that may be playing important roles in the years to come are described.     

Viral diseases in commercially exploited crabs: a review

Viruses and viral diseases of crabs were observed and investigated earlier than the first observation of viruses in shrimp. In fact, crabs were used as biological models to investigate crustacean virology at the beginning of shrimp aquaculture development. More than 30 viruses have been reported in crabs, including those related to the known virus families Reoviridae, Bunyaviridae, Roniviridae and a group of Bacilliform enveloped nuclear viruses. This review reports data on several important viral diseases of crabs, particularly those associated with pathology of organs and tissues of commercially and ecologically significant host species.     

环境病毒的宿主鉴定技术进展

病毒是地球上丰度最高的微小生命粒子,通过调控宿主的群落结构、介导宿主死亡和参与水平基因转移等方式影响着生物地球化学循环和地球生命演化。近年来,宏基因组学的发展实现了在全球尺度上对环境病毒的大规模探索和研究,大量新的病毒基因组被发掘,病毒在全球生态过程和生物地球化学循环中的角色和贡献也得到进一步认知。病毒在环境中的重要作用是通过感染宿主实现的。然而,环境病毒的宿主鉴定工作远落后于环境病毒基因组测序研究。本文综述了目前病毒宿主鉴定的主要技术及其优缺点和应用场景,总结了病毒的宿主鉴定在病毒生态学研究和生物工程领域的重要价值,并初步展望了未来病毒宿主鉴定技术的发展方向。     

西番莲花叶病研究及防治方法的进展

西番莲Passiflora edulis是热带亚热带重要水果之一。病毒病害能造成西番莲叶片花叶、褪绿，果实畸形、木质化以及植株矮化，严重危害西番莲种植、制约增产。花叶病是危害西番莲的主要病毒病之一，引起西番莲花叶病的主要病毒有：黄瓜花叶病毒属Cucumovirus病毒、马铃薯Y病毒属Potyvirus病毒、双生病毒属Geminivirus病毒等。本文综述了西番莲花叶病分布及症状特征，病毒生物学特性、传播途径以及检测技术，为研究其发病机制提供参考，同时提出西番莲花叶病害综合防治方法。     

Bioinformatics databases and tools in virology research: an overview

Viruses are major factors of human infectious diseases. Understanding of the structure-function correlation in viruses is important for the identification of potential anti-viral inhibitors and vaccine targets. In virology research, virus-related databases and bioinformatic analysis tools are essential for discerning relationships within complex datasets about viruses and host-virus interactions. Bioinformatic analyses on viruses include the identification of open reading frames, gene prediction, homology searching, sequence alignment, and motif and epitope recognition. The predictions of features such as transmembrane domains, glycosylation sites, and protein secondary and tertiary structure are important for analyzing the structure-function relationship of proteins encoded in viral genomes. Biochemical pathway analysis can help elucidate information at the biological systems level. Microarray analysis provides methods for high throughput screening and gene expression profiling. Virus-related bioinformatics databases include those concerned with viral sequences, taxonomy, homologous protein families, structures, or dedicated to specific viruses such as influenza and herpes simplex virus (HSV). This review provides a guide and overview of computational programs for these analyses as a resource for genomics and proteomics studies in virology research. These resources are useful for understanding viral diseases, as well as for the design and development of anti-viral agents.     

Biological roles and functional mechanisms of arenavirus z protein in viral replication

Arenaviruses can cause severe hemorrhagic fever diseases in humans, with limited prophylactic or therapeutic measures. A small RING-domain viral protein Z has been shown to mediate the formation of virus-like particles and to inhibit viral RNA synthesis, although its biological roles in an infectious viral life cycle have not been directly addressed. By taking advantage of the available reverse genetics system for a model arenavirus, Pichinde virus (PICV), we provide the direct evidence for the essential biological roles of the Z protein's conserved residues, including the G2 myristylation site, the conserved C and H residues of RING domain, and the poorly characterized C-terminal L79 and P80 residues. Dicodon substitutions within the late (L) domain (PSAPPYEP) of the PICV Z protein, although producing viable mutant viruses, have significantly reduced virus growth, a finding suggestive of an important role for the intact L domain in viral replication. Further structure-function analyses of both PICV and Lassa fever virus Z proteins suggest that arenavirus Z proteins have similar molecular mechanisms in mediating their multiple functions, with some interesting variations, such as the role of the G2 residue in blocking viral RNA synthesis. In summary, our studies have characterized the biological roles of the Z protein in an infectious arenavirus system and have shed important light on the distinct functions of its domains in virus budding and viral RNA regulation, the knowledge of which may lead to the development of novel antiviral drugs.     

The search for viruses in bovine semen, a review

Viruses reported in bovine semen include those of foot and mouth disease, bluetongue, bovine leukemia, infectious bovine rhinotracheitis, bovine viral diarrhea, ephemeral fever, and lumpyskin disease. Bovine enteroviruses, a parapoxvirus (paravaccinia), and several uncharacterized viruses have also been isolated. Their presence in semen has been recognized by a variety of animal inoculation and cell culture techniques. The prevalence of viruses in semen and the attendant impact on fertility is largely unknown. The widespread distribution of frozen semen, an ideal system for preserving viral infectivity, indicates this product could serve as an important vehicle for transmission of viruses to uninfected herds or areas. This concern has placed restrictions on international movement of semen. Improved methods for detecting viruses in semen are necessary to assess the importance of the problem and to develop realistic precautionary measures.     

媒介昆虫-病毒-植物互作对生物入侵的影响

媒介昆虫-病毒-植物互作关系复杂多样。虽然相关的研究较多, 然而有关三者互作对于生物入侵的影响还知之甚少。已有证据表明, 寄主植物对病毒的敏感性和对媒介昆虫的适合性、媒介昆虫对寄主的适应能力等因素影响三者互作关系。当寄主植物易感病并且对媒介昆虫的适合性低, 而媒介昆虫对寄主植物的适应能力强时, 媒介昆虫与植物病毒之间很可能建立间接互惠关系, 这种互惠可促进媒介昆虫入侵和病毒病流行。此外, 媒介昆虫与植物病毒之间中性或偏害的互作关系对于外来生物入侵的促进作用也不容忽视。鉴于三者互作对于生物入侵的重要性, 今后需要对不同物种所组成的多种组合进行比较研究, 并采用多种方法揭示互作的生理和分子机制。     

Les virus des eaux usées et leur élimination au cours des traitements des effluents pollués

Viruses are a biological pollution in wastewaters. Their elimination results from treatment processes of polluted effluents in the sewage treatment plant or in the lagooning pilot plant. Bacteriophages are regarded as viruses models and as indicators of faecal contamination. In an expiremental study, two pilot plants laboratories have been used (natural lagooning and activated sludge pilot) to follow and evaluate the elimination rate of somatic coliphage X-174 or the ARN F-specific one MS2. The two phages removal, ranging to 99 % in the two systems, are related with previous values on enteric viruses. The main factors of viral removal and inactivation (adsorption to solid matters, microbial activities, solar radiations, dissolved matters) have been studied too. Adsorption of infectious particles to solid matters is a very significant process. This was obtained in the pilot plant laboratory experiment and in in vitro study (X-174 or MS2 in the presence of the kaolinite or the montmorillonite). Otherwise, 12 hours of solar rays exposition were efficient ro inactivate nearly the whole phagic suspension. The ciliate protozoan Tetrahymena pyriformis does not seem to have an important role for elimination of phages (X-174 and MS2.     

RNA viruses in the sea

Viruses are ubiquitous in the sea and appear to outnumber all other forms of marine life by at least an order of magnitude. Through selective infection, viruses influence nutrient cycling, community structure, and evolution in the ocean. Over the past 20 years we have learned a great deal about the diversity and ecology of the viruses that constitute the marine virioplankton, but until recently the emphasis has been on DNA viruses. Along with expanding knowledge about RNA viruses that infect important marine animals, recent isolations of RNA viruses that infect single-celled eukaryotes and molecular analyses of the RNA virioplankton have revealed that marine RNA viruses are novel, widespread, and genetically diverse. Discoveries in marine RNA virology are broadening our understanding of the biology, ecology, and evolution of viruses, and the epidemiology of viral diseases, but there is still much that we need to learn about the ecology and diversity of RNA viruses before we can fully appreciate their contributions to the dynamics of marine ecosystems. As a step toward making sense of how RNA viruses contribute to the extraordinary viral diversity in the sea, we summarize in this review what is currently known about RNA viruses that infect marine organisms.