Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/53427
標題: 開發植物及其產品檢疫技術
Development of Quarantine Techniques for Plants and Plant Products
作者: 曾國欽, 陳啟予,陳煜焜,陳隆鐘,張雅君
關鍵字: 應用研究;Seed-borne Disease;植物保護類;聚合酵素連鎖反應;十字花科黑腐病菌;種傳病毒;檢測;萵苣嵌紋病毒;長喙殼菌;鑑定;木材檢疫;煙草脆葉病毒;番茄輪點病毒;山芥菜嵌紋病毒;反轉錄聚合酶連鎖反應;土傳病害;分子檢測;Polymerase Chain Reaction;Xanthomonas campestris pv. campestris;Seed-borne Viruses;Detection;Lettuce Mosaic Virus;Ophiostoma;Identification;Wood Quarantine;Tobacco Rattle Virus;Tomato Ringspot Virus;Arabis Mosaic Virus;RT-PCR;Soil Borne Disease;Molecular Detection
摘要: 
1. Development of detection techniques for plant improtant quarantine pests:The objective of this study is to develop a rapid detection technique for seed-borne bacteria. We will detect Xanthomonas campestris pv. campestris (Xcc) in natural infected seeds from the field by the PCR based detection technique developed in the previous study. The factors affecting senstivity of detection of Xcc will be studied. The standard operating procedures (SOPs) will be developed and used for seed quarantine. 2. Development of quarantine techniques for seedling and the material of agriculture production: In order to construct a platform which is design to screen the seedling and the material of agriculture production, the identification with quality of seedling species will be supported by molecule health supervision and the platform of nucleic acid quantification system. Further more, the connection of data side servicing base and the line transporting of disease event report will be used to predict and assess to realize the process of disease and the possibility of happening immediately. Therefore, the technology which is built up will be adapted to address the work of quarantine and to pass the ISO of national community. Besides, super sensitive tech of detecting the sequence will be allowed to be used to pursuit the information and evident of research. On the other hand, by exchange quantified data from PAGE and the developing the commercial good for those who generate the vegetable and seeding will be a good looking in the future.3.The development of the tools and techniques for detecting seed-borne viruses of economically important vegetables:Plant viruses can be transmitted by their hosts' seeds. This common phenomenum makes plant seed an inspection target of quranteen. This project will develop detection tools and techniques for the inspection scheme of some important seed-borne viruses of vegetables, such as Lettuce mosaic virus of lettuce. The virus will be isolated and purified, the viral CP gene will be cloned and sequenced, and the specific antiserum as well as molecular probes will be produced. The produced tools, i.e. antiserum, probes, primers, etc, and the developed techniques will provide sensitive and reliable ways of detection for the inspection scheme of vegetable seed-borne viruses.4. Development of detection system for important plant viruses of solanaceous seeds:Tobacco rattle virus (TRV), Tomato ringspot virus (ToRSV) and Arabis mosaic virus (ArMV) which are important quarantine viruses, can infect vegetable seeds of the family Solanaceae. To prevent these viruses from being introduced into Taiwan through imported seeds, this project is going to develop the specific primers of TRV, ToRSV and ArMV as well as antisera of TRV and ArMV, and thus establish a detection system for important seedborne viruses. The following works will be executed this year. (1) Test the specificity of TRV primers with infected tissues, and establish the best RT-PCR reaction condition. (2) Develop multiplex RT-PCR detection method for TRV and ArMV. (3) Mix the seeds with TRV and ArMV extracts to mimic infected seeds and then check the seeds by multiplex RT-PCR. (4) Investigate imported seeds for TRV and ArMV.5.Exploitation of techniques for detecting harmful organisms on imported wood:The members of Ophiostoma commonly occur on woody plants, adversely affecting the wood quality. Some species of Ophiostoma are particularly the causal agent of serious forest diseases. Woods required for markets are largely from importation. It underlies the risk for the non-indigenous Ophiostoma fungi to be introduced into Taiwan. The aim of this project is to establish efficient detection and identification approaches for Ophiostoma species. The fungi will be constantly isolated from various imported wood materials. Microscopic observation, cultivation, DNA extraction and sequencing will be carried out for the acquired isolates. In the combination of morphology, biological features, mating type and DNA sequences, The method of species identification of Ophiostoma species can be obtained. Furthermore, the gathering of those data for different Ophiostoma species from imported wood will form the basis to develop rapid detection and identification techniques.

一、 重要植物檢疫有害生物檢測技術之研發(中興大學曾國欽):本研究擬針對重要之種媒細菌開發快速檢測之標準流程,本研究將利用前期所開發之十字花科黑腐病菌檢測技術進行田間實際檢測,並對相關影響因子進行探討,本研究所開發出之十字花科黑腐病菌檢測標準流程將提供防檢疫相關人員檢測十字花科黑腐病菌之用。 二、 開發蔬菜及其使用資材檢疫技術(中興大學陳隆鐘):本計畫擬針對種苗及使用資材來作檢測,應用分子標記技術進行種子品種的純度鑑定,並同時進行種子健康分子檢測的工作,以核酸定量系統為平台,結合基因資料庫,並配合各種病害發病門檻的資料,將可在短期內預測病害發生或進展的可能性,進而可應用於檢防疫工作,以期使此檢測技術符合國際規範標準作業程序之訂定,並得到國際標準之認證,期與國際間各國檢疫接軌。此外,高靈敏度技術亦可用於特定核酸序列的定量追蹤研究上,以量化資料取代電泳條帶分析,或是開發為商品化的檢測套組供政府相關單位及蔬菜種苗業者使用。 三、 重要茄科蔬菜種子病毒標準檢測流程之建立(台灣大學張雅君):以能感染茄科蔬菜種子的菸草脆葉病毒(Tobacco rattle virus, TRV)、番茄輪點病毒(Tomato ringspot virus, ToRSV)和山芥菜嵌紋病毒(Arabis mosaic virus, ArMV)為對象,研發其專一性引子對和RT-PCR檢測法,並製備TRV及ArMV的專一性抗血清,以建立進口種子重要檢疫病毒之檢測系統。今年將執行下列重要工作: (1) 以感染TRV之植物材料測試TRV引子對之專一性,同時建立最佳的RT-PCR反應條件。 (2) 嘗試將TRV和ArMV的引子對配合mt引子對,建立multiplex RT-PCR的反應條件。 (3) 將TRV和ArMV的材料與茄科蔬菜種子混合,再以我們所建立的multiplex RT-PCR法進行種子病毒檢測。 (4) 同時監測國外進口種子TRV和ArMV的存在情形。 四、 重要蔬菜種傳病毒檢測工具與技術之開發:病毒藉由寄主的種子傳播是一個普遍的現象,因而造成種子在進出口時常成為檢疫的對象。本計畫將針對數種重要的蔬菜種傳病毒開發種子檢測工具和檢測技術,以利種子檢疫工作之遂行。本年工作以菊科蔬菜之種傳病毒(如lettuce mosaic virus)為主要檢測對象。工作項目包括病毒之分離與純化、病毒外鞘蛋白基因之選殖與定序、病毒抗血清之製備與血清學試驗等預期將可產出專一性抗血清、分子探針與引子對,並發展包括ELISA, RT-PCR, IC-RT-PCR等技術,供種子檢測之用。 五、 輸入木材有害生物檢測技術之研發:長喙殼菌屬之真菌普遍存在於林木中,影響木材之品質,有些種類更是世界上重要之林木病害,我國林木原料大多仰賴國外進口,因而存在著外來長喙殼菌侵入之風險。本研究之目的即在建立有效偵測及鑑定進口木材中長喙殼菌之技術,藉由持續的從不同進口材質中分離菌株,進行觀察、培養、與DNA萃取及解序,建立形態鑑定之觀察標準、生長條件之判別標準與配對測試菌株,更進一步將所收集菌株之rDNA片段進行解序,與基因庫中存有之種類進行序列比對、及親緣關係分析,來進行鑑定之推衍。實驗將累積不同種類之形態、生理、配對、分子資料,建立進口木材中潛在有害長喙殼菌之資料庫,並以此資料庫為基礎,發展快速之檢測方法。
URI: http://hdl.handle.net/11455/53427
其他識別: 96農科-14.4.1-檢-B1
Appears in Collections:生命科學系所

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