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Broad-Spectrum and Specific Tools for Diagnosis of Different Serogroups of Thrips-Borne Tospoviruses
|關鍵字:||技術發展;Tospovirus;植物保護類;番茄斑萎病毒;病毒鑑定;血清製備;核酸選殖;核酸探針;Virus Identification;Antiserum Preparation;cDNA Cloning;Nucleic Acid Probe||摘要:||
番茄斑萎病毒屬(Tospovirus)為一群帶有外套膜並可經薊馬傳播的病毒,在世界各地造成許多重要經濟作物重大損失,此病毒寄主範圍廣大,目前已有記載的達82科800餘種植物,包含茄科作物、葫蘆科作物、菊科作物、豆科作物等,並已成為此類作物生產限制因子之一.由於此病毒的寄主範圍非常廣泛,且病徵差異大,目前以血清類緣關係及核鞘蛋白(nucleocapsid protein, NP)核酸序列分析為主要分類依據.目前蕃茄斑萎病毒屬有十個不同的血清型(serogroup),最主要的四個血清型是血清型I、II、III及IV.第一血清型(serogroup I)以蕃茄斑萎病毒(TSWV)為代表,第二血清型(serogroup II)含花生輪點病毒(GRSV)及蕃茄黃化斑點病毒(TCSV).第三血清型(serogroup III)以鳳仙花壞疽斑點病毒(INSV)為代表,第四血清型(serogroup Ⅳ)含西瓜銀斑病毒(WSMV)及花生頂芽壞疽病毒(PBNV).由於此病毒顆粒具有套膜,粒子極不穩定,所以純化困難,抗血清製備不易,因此如何在核酸分子層次上研發病毒的檢測工具實為當務之急.此三年的研究計畫主要目的在開發蕃茄斑萎病毒屬主要血清型廣效性及專一性診斷之工具,包括 1) 研發簡併性(degenerate)引子作為廣效性檢測不同血清型之tospovirus病毒之工具; 2) 研發專一性單一血清型之病毒核酸探針; 3) 構築對特定血清型具有抗性的轉基因菸草來做生物分析,檢測各個不同血清型的tospovirus病毒; 4) 研發廣效性能夠檢測不同血清型tospovirus之抗血清,及專一檢測單一血清型之病毒抗血清.經由本計畫之研發,我們將能對台灣及全世界在田間tospovirus病害上提供簡易且精確快速之診斷工具.
The genus Tospovirus encompasses a group of enveloped plant viruses that cause great losses in many economically important crops worldwide and becomes to be the growth-limiting factor of many crops. Tospoviruses have a wide host range and are known to infect more than 800 different plant species within 82 families including Solanaceae, Cucurbitaceae, Compositae, Leguminosae. Tospoviruses are transmitted by thrips. Since the very broad host range and the highly varied symptomology found within the genus, the serological relationships and sequence homology of the nucleocapsid (N) protein gene are important criteria for classifying the taxonomic position of a virus in the genus Tospovirus. Based on serological cross-reactivity and sequence homology of the N gene, ten serogroups have been recognized in the Tospovirus genus. The major serogroups are serogroup I (tomato spotted wilt virus, TSWV), serogroup II (groundnut ringspot virus, GRSV; tomato chlorotic spot virus, TCSV), serogroup III (impatiens necrotic spot virus, INSV) and serogroup IV (watermelon silver mottle virus, WSMV; peanut bud necrosis virus, PBNV). Since the enveloped particles of tospoviruses are unstable and rapidly break down once outside the host tissue, it is difficult to prepare sufficient amount of whole virus particles suitable for immunization. Thus, develop approaches based on molecular biology techniques for the detection of tospoviruses become very important. The main objectives of this three-year proposal are to develop broad-spectrum and specific tools for diagnosis of different serogroups of thrips-borne tospoviruses. We will 1) design degenerated primers for broad-spectrum identification and diagnosis of different serogroups of tospoviruses; 2) develop nucleic acid probes or primers for individual serogroup of tospoviruses; 3) establish a simple and fast approach to detect and differentiate tospoviruses using transgenic tobacco with specific resistance to TSWV, INSV, GRSV, and WSMV; 4) generate antisera which can broad-spectrum and specifically react with different serogroups of tospoviruses. Though these efforts, we will be able to provide easy and reliable tools for detection and diagnosis of different serogroups of tospoviruses in Taiwan and worldwide.
|Appears in Collections:||植物病理學系|
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