Identification of plant-parasitic nematodes by PCR amplification of DNA fragments1

A study concerning the detection and characterization of a DNA fragment from plant-parasitic nematodes to be used as a molecular marker for the identification of different nematodes is described. A fragment of DNA, which is known to consist of a variable region flanked by two conserved regions, has been studied by using PCR amplification. A portion of about 600 nucleotides at the 5’end of the larger rRNA gene has been amplified in different nematodes, using heterologous primers which hybridize with the conserved regions. The results obtained clearly indicate that the same primers can be used for the amplification of this segment in nematodes of different species and of different genera: Meloidogyne artiellia, M. incognita, Xiphinema index, X. diversicaudatum and Globodera pallida. The amplified regions have been partially sequenced. The nucleotide sequences have been analysed by comparison with the published sequence of Caenorhabditis elegans.     

Real-time PCR,a great tool for fast identification,sensitive detection and quantification of important plant-parasitic nematodes

Plant-parasitic nematodes can cause significant damage to agricultural crops and forests worldwide, resulting in major economic losses. Some nematode species do not occur in all areas and are regulated as quarantine organisms. To avoid introduction and spread of these organisms, fast, simple and reliable detection and identification methods are needed, that help plant diagnostic services such as reference centres or national plant protection organizations (NPPOs) to rapidly identify suspicious nematodes. Real-time PCR is one of the fastest, most sensitive and reliable methods to fulfil this task. It is a DNA-based method that is easy to learn with the only requirement of having a specific thermocycler (Real-time Platform) and the appropriate chemistry. Real-time PCR provides very sensitive detection and species-specific identification with the potential to quantify target organisms if required. Following DNA extraction, results can be seen in 1–3 h and management decisions applied. Real-time PCR can be used for high-throughput analysis of many samples and in some cases for multiplexing, allowing for identification of more than one species in a single reaction. Over the past 15 years, real-time PCR methods have been developed for the main plant-parasitic nematodes, in particular the regulated species. This paper reviews the achievements in plant nematology diagnostics using real-time PCR as the method of choice for fast and reliable detection, identification and even quantification of plant parasitic nematodes.     

美国检疫线虫及潜在检疫线虫风险分析

近些年来，随着国际间贸易往来的逐渐增加，携带线虫入境的机会越来越大。因此，对入境线虫进行风险分析及管理显得越来越必要和急需。我国对检疫线虫的风险分析报道较少，对潜在检疫线虫的风险分析尚未见报道。本文简单介绍了美国对检疫线虫及潜在检疫线虫风险分析的方法，以供我国学者和同行以及有关决策部门参考。     

‘99昆明世博会参展植物寄生线虫的检疫初报

本次博览会参展植物种类多,来源广,带土普遍疫情复杂。在严处国内外的参展植物中截获矛线目和垫刃目的１３个科的５６个属,其中在参展植物中截获我国对外检疫潜在危险性线。     

Viruses transmitted by nematodes1

The transmission of plant viruses by nematodes is remarkable in involving only two distinct groups of viruses, nepo viruses and tobraviruses, and being limited to longidorid, Paratrichodorus and Trichodorus nematodes respectively. Tobraviruses and their associated vector nematodes are not discussed here. Only 11 of the 36 described nepoviruses are transmitted by nematodes, and 6 of these 11 viruses are present in Europe naturally associated with 8 virus-vector longidorids. Specific relationships exist between the serologically distinct viruses and their vector nematode species. Specificity is largely determined by the virus coat protein and by an inherited ability of the nematode to retain virus particles at specific sites within its oesophagus. This specific relationship can be quite subtle, extending to populations of vector nematodes and also to virus isolates which apparently are serologically indistinguishable. Several serological and/or symptomatological variants of nepoviruses may be present at a field site in association with one or more vector nematode species. The exposure of different crops and new cultivars to these virus and vector combinations will probably result in the occurrence of further nematode-transmitted virus diseases. New methods for suppressing damage to crops caused by these diseases are required including the likely use of transgenic resistant cultivars.     

北美墨天牛属害虫的检疫鉴定

墨天牛属ＭｏｎｏｃｈａｍｕｓＧｕｅｒ害虫因传播松树毁灭性有害生物松材线虫Ｂｕｒｓａｐｈｅｌｅｎｃｈｕｓｘｙｌｏｐｈｉｌｕｓ而受到广泛的关注。北美约有8种墨天牛,主要危害松属树木,少数种类还危害冷杉、云杉及黄杉属树木,而这些树木又是制作木质包装的主要材料,极易随     

检疫性蛾类鉴定方法和标准的研究进展

由于检疫性蛾类多数种类个体小,形态难以观察,且与其他蛾类害虫在形态上具有一定的相似性,区别和鉴别难度大,造成该类检疫害虫传播扩散的风险加剧。一方面,该类害虫的幼虫啃食水果、木材等,使其品质下降,严重影响水果、木材等进出口贸易;另一方面,检疫害虫的扩散也对农林业生产构成了巨大威胁。因此,蛾类害虫的精准鉴定已成为国内外普遍关注的问题。该文对我国检疫性蛾类的种类进行系统性汇总,并对鉴定方法及鉴定标准等方面的研究进展进行综述,分析目前存在的问题和不足,并结合技术创新以及口岸实际检疫需求,提出今后的发展方向,以期为检疫性蛾类新型精准鉴定技术研发及其入侵防控策略制订提供参考。     

Transmission of tobraviruses by trichodorid nematodes1

Transmission of tobacco rattle tobravirus by Paratrichodorus and Trichodorus has been known since 1961. However many of the studies of these virus-vector associations have been uncritical. Here we describe a system to study acquisition and transmission of individual tobravirus isolates by given species of Paratrichodorus and Trichodorus. Results from initial experiments in which this system was used suggest that different isolates of tobacco rattle tobravirus differ in vector specificity.     

全国口岸进境邮寄物中截获线虫的疫情分析

本文对全国口岸2003~2016年4月从邮寄物中截获的线虫进行了汇总与分析,归纳了邮寄物中截获寄生线虫的种类、来源等,结果表明,全国口岸从邮寄物中共计截获线虫42属、4 448批次,其中包括检疫性线虫11种、49批次,随着邮寄物品数量和种类的增加,邮检中线虫的截获率逐年增加。目前进境邮寄物中线虫的疫情十分复杂,应对进境苗木、植物产品、蔬菜水果等加强检疫监管,以防止线虫疫情进一步扩展。     

我国进境植物检疫性菌物检疫鉴定标准浅析

我国现行进境植物检疫性有害生物名录中共收录130种检疫性菌物.本研究在收集整理现行检疫性菌物检疫鉴定标准基础上,对该类标准的历史、现状进行了分析,探讨了检疫性菌物标准的特点,针对我国植物检疫性菌物检疫鉴定标准的发展与完善提出了建议.     

Image analysis for the identification of the quarantine pest Tilletia indica*

Diagnosis of the teliospores of Tilletia indica (Karnal bunt) detected in wash tests is complicated by the fact that wheat grain can be contaminated by ‘look‐alike’ species, namely Tilletia walkeri (ryegrass bunt) and Tilletia horrida (rice smut). Although morphological diagnosis is possible when there are relatively large numbers of teliospores present, it is difficult with only a few spores as there are significant overlaps in the characteristics of each species. Molecular methods can confirm presumptive identifications, but these take 2–3 weeks. Image analysis offers the potential for more rapid confirmation. An image analysis system was developed for use on bleached spores. Bleaching spores reveals additional morphological characters (spore profile and spore wall layers) that may be used in an image analysis system to discriminate species. The image‐processing software automatically locates spores on a given image and calculates perimeter, surface area, number of spines and spine size, maximum and minimum ray radius, aspect ratio and roundness. Principal components analysis (PCA) is performed on the parameters to obtain a linear separation of spore species. Accuracy of 97% in separating T. indica and T. walkeri has been achieved in preliminary tests using PCA, but further evaluation is required.     

植物线虫分子鉴定研究进展

植物线虫可危害农作物和林木,对它们的准确鉴定是防治植物线虫病害的基础。由于植物线虫很小,而且在形态上种间常有覆盖,而种内有较大的变异,仅依据形态特征很难鉴定。分子鉴定技术给植物线虫的检测和鉴定提供了快速、精确、可靠的方法。文章综述了植物线虫分子的DNA提取、分子鉴定靶标序列的选择及分子鉴定方法等方面的研究进展及现状,以促进对植物线虫分子鉴定更深入的研究。     

基于多重PCR技术的高通量测序在植食性昆虫食谱鉴定中的潜在应用

多重PCR技术因成本低、灵敏度高、快捷、高效等优点而被广泛用于遗传病鉴定和传染病传播、食品掺假、污染鉴定以及病虫害检测中,但多重PCR技术在动植物分类中应用较少,在植食性昆虫食谱鉴定中应用更少。该文详细介绍了植食性昆虫食谱分子鉴别技术,多重PCR技术要素、种类和应用以及与其他技术的结合应用,高通量测序中多重PCR捕获技术,并对多重PCR技术与高通量测序技术的结合在动物食谱鉴定中的潜在应用进行了展望。     

PCR法检疫鉴定番石榴实蝇

利用mtDNA序列,设计出两对特异性引物,应用定性PCR技术检疫鉴定番石榴实蝇.本方法适用于成虫鉴定和未成熟虫态(卵、幼虫、蛹)的快速检疫鉴定.     

金钱松实时荧光PCR鉴定方法的建立

本研究根据金钱松核糖体基因的内转录间隔区(ITS)设计特异性引物与荧光探针,建立了金钱松的实时荧光PCR检测方法.利用该方法检测4个不同来源的金钱松样本,均能够得到典型的扩增曲线,与53个松柏科其他树种如日本五针松、罗汉松、湿地松样本无交叉反应,表明了该方法的特异性.灵敏度可达1.6×10-5ng/μL.该方法可快速、...     

中国检疫性有害生物信息管理与辅助鉴定系统的研究(一)

入世为中国植物检疫带来机遇和挑战，计算机技术等高新技术的应用对我国全面实施《SPS协定》起着重要的作用。本研究以软件工程原理知专家系统技术为基础，采用LUBAN模型和Delphi编程语言，针对中国检疫性有害生物信患管理与辅助鉴定系统(QPⅡS)进行了研制。本文主要介绍了QPⅡS的研究目的、方法、系统定义和设计。     

专家系统与检疫性有害生物的辅助鉴定

专家系统属计算机人工智能技术范畴，能够解决植物检疫工作中检疫性有害生物的辅助鉴定问题。本文在介绍专家系统基本结构和类型基础上，分析了植物检疫领域专家系统的应用，并对检疫性有害生物辅助鉴定专家系统的研究方法进行了初步探讨。     

A biotechnological strategy involving monoclonal antibodies for improvement of potato farming by identification and quantification of potato cyst nematodes in soil samples1

The two closely related nematode species Globodera rostochiensis and G. pallida are one of the major problems encountered in potato cultivation. There is a spectrum of potato plant genes known, which confer resistance to these species and their pathotypes. Potato growing in The Netherlands has to follow strict rules to control spread of the pests. Since distinction between the two nematode species is difficult, a rapid and reliable identification method is needed to allow better use of existing and forthcoming resistant potato cultivars. The aims of this project were: (1) identification and partial purification of species-specific proteins from the nematodes, (2) production of species-specific monoclonal antibodies, and (3) development of a screening test for qualitative and quantitative determination of Globodera spp. in soil samples.