| 摘要： 一、輸出入植物檢疫有害生物之診斷鑑定及檢疫處理技術研發(一)邊境查驗檢出檢疫有害生物之整合鑑定及分析系統1. 加強進出口農產品檢疫鑑定作業分層負責系統。2. 開辦重要檢疫害物之蟎類及蚜蟲鑑定技術訓練。3. 協助緊急檢疫樣品鑑定。(二)建立檢疫重要性果實蠅診斷鑑定技術、疫情資訊及教育訓練系統1. 強化果實蠅鑑定能力，提供診斷所需資訊，提供教育訓練、諮詢及果實蠅分類鑑定服務等。2. 建立及修訂國內外具檢疫重要性果實蠅鑑定圖譜。3. 整理並發表臺灣本地果實蠅害蟲名錄。(三)利用DNA條碼建立纓翅目、鱗翅目、雙翅目及半翅目等檢疫有害生物分子鑑定技術建立『中華民國輸入植物或植物產品檢疫規定』所列纓翅目、鱗翅目、雙翅目、鞘翅目等百餘種之各目檢疫害蟲DNA條碼。另按我國農業資料庫之作物、花卉、蔬果所列害蟲資料，建立纓翅目、鱗翅目、雙翅目、半翅目及其它各目重要害蟲DNA條碼。除了針對甲乙類重要檢疫害蟲開發快速檢測試劑外，另應用DNA資料設計探針開發重要花薊馬生物晶片鑑定系統。(四)小果番茄冷藏檢疫處理技術小果番茄果實接種標的果實蠅之小規模冷藏殺蟲試驗:依低溫抗性比較試驗結果，以抗冷性最強之果實蠅及齡期作為殺蟲試驗之供試蟲期，即果實內之幼蟲達到最適齡期時，再放入冷藏庫進行試驗，探討低溫檢疫處理條件 (五)研發輻射照射應用(五)於番石榴鮮果實檢疫處理技術利用國內現有輻照處理設施進行番石榴鮮果檢疫照射處理試驗，據以草擬一套適用於外銷番石榴輻射照射檢疫處理標準流程，提供防檢局做為相關檢疫處理方法之參考。 二、重要檢疫及新興有害生物風險評估及管制措施研究(一)我國重要檢疫病毒類有害生物之國際疫情分析及檢測技術開發為維持我國有害生物清單中病毒類資料之準確性，我們將繼續檢視並修正病毒清單中剩餘的169筆資料之學名與分類，同時增列重要植物檢疫病毒至有害生物清單。重新彙整有害生物清單中共475筆病毒類的修正資料與所收集的相關文獻。此外，對近年來受到多國重視可感染番茄和番椒之新種傳病毒—番茄斑駁嵌紋病毒(Tomato mottle mosaic virus, ToMMV)，蒐集其文獻和病毒序列資料，設計 ToMMV 專一性引子對，研發RT-PCR檢測方法。將這些完整的病毒類修正資料，以及所研發之 ToMMV 檢測方法，提供給植物檢疫單位參考使用。(二)具潛在檢疫風險線蟲評估鑑別技術開發因應進口無土栽培介質及植株附帶栽培介質中檢出之重要植物檢疫線蟲有比例逐漸增加之趨勢，且為求有效預防高風險性外來種之入侵與散佈，本計畫將透過國際線蟲研究資料、進口植物與其相關產品中所攔截之線蟲，進行線蟲資訊之蒐集與分析，優化現有之葉芽線蟲屬內種間之PCR快速分子檢測技術，逐步建立無土栽培介質之線蟲群相資料庫，以及建立重要檢疫線蟲之偵測鑑定於實務場所之作業流程。(三)亞洲臨近地區新興鱗翅目害蟲之風險評估與鑑識秋行軍蟲(Spodoptera frugigperda，英名縮寫為FAW)為近年全球雜糧作物之重大害蟲。由於這種昆蟲快速地在2019年6月入侵我國，為了即早建立防治體系所需要之基礎知識，我們將利用生態棲位模擬(ecological niche modeling)與預測物種分布(predicted species distribution model)分析技術，推測此種蛾類在臺灣本島未來的擴張範圍。釐清秋行軍蟲與同屬物種與不同屬但形態上易混淆物種，以及所有在臺灣的作物上可出現的夜蛾總科昆蟲之鑑識方法，並藉由比較博物館標本典藏瞭解各地秋行軍蟲之表型差異。採集因應分子親緣關係與族群遺傳學所需之樣本，以利在未來持續監測臺灣島內族群遺傳結構之動態、抗藥性相關遺傳特質的變動，以及入侵來源。我們希望這個計畫的成果除了能夠協助防治之外，還能夠培訓新興有害生物之鑑識與診斷人才，發表相關科學成果，並提供政府、農民與試驗單位鑑識服務。 三、國際植物有害生物疫情研析及工具開發(一)植物檢疫有害生物資料庫及核准輸入植物清單與檢疫條件查詢系統維護及功能擴充植物檢疫規定是業者自國外進口農產品的重要參考資料，此規定旨在避免國外植物疫病蟲害侵入，並制止其蔓延。本計畫在已建置之植物檢疫有害生物疫情資料庫框架基礎上，進行三項工作: (1) 維護並更新「植物檢疫有害生物疫情資料庫」之有害生物及植物基本資料（含學名及分類地位）; (2)新增有害生物風險評估簡表生成功能; (3) 設計植物疫情資訊之科技文獻自動化知識擷取機制。本計畫除持續維護、擴充”核准輸入植物清單與檢疫條件查詢系統”與”檢疫有害生物資料庫”之功能。(二)蘭花病蟲害疫情資料蒐整及整合性防治技術開發 臺灣外銷蘭花主要有蝴蝶蘭、蕙蘭與嘉德麗雅蘭等，在面對國際化的出口競爭壓力中，在出口檢疫時，病蟲害的發生，特別是害蟲的存在，往往導致出口障礙與出貨品質不佳。本研究擬針對蝴蝶蘭、蕙蘭與嘉德麗雅蘭常見病蟲害，從清潔種苗制度、園區病蟲害管理作為與病蟲害發生時的防治作法，建立病蟲害的整合式防治技術。 四、重要作物病蟲害診斷、檢測鑑定技術之研發及應用(一)農業生態系統之金花蟲亞科: 分類、食性及氣候變遷下之族群變動金花蟲科(Chrysomelidae)為為鞘翅目(Coleoptera)中種類最多樣的一個科，台灣已記錄的種類超過六百種；往昔的研究都只著重於新種及新紀錄的發表，且採集地點相當零散。本研究為現代化分類學研究的開端，針對金花蟲亞科Chrysomelinae 作分類修訂，以全面性及定期性的方式採集標本，探討屬級的分類地位，詳細研究每個種類的診斷特徵、成蟲出現時期、寄主植物及地理分布，並重新檢查已知種的模式標本，以建立一個扎實且有系統性的診斷鑑定研究。(二)番茄系統性病害之鑑定與防治技術開發番茄為台灣重要果菜類蔬菜，系統性的病害常由病毒與細菌引起，造成嚴重損失。目前主要的病毒病害為雙生病毒感染引起的番茄捲葉病與黃化捲葉病，主要以抗（耐）品種及媒介昆蟲粉蝨為主要防治對策，但調查發現雙生病毒在番茄上常見複合感染或基因重組，使得病毒變異大，引進抗病基因的番茄品種抗病效果逐年遞減。因粉蝨已適應台灣自然環境且雜食性強，要以藥劑控制粉蝨達到防治此病害的效果必須密集施藥，抗耐病品種培育仍是最有效且環保的方法，目前抗耐病品種篩選仍以置於溫網室中利用粉蝨媒介感染，篩選結果因粉蝨族群及傳毒效率而異，使實驗變數增大。而在細菌病害上，台灣田間可在番茄維管束蔓延的病原菌主要有青枯病菌(Ralstonia solanacearum)及莖腐病(Pectobacterium carotovorum subsp. carotovorum)。近年田間零星出現新興髓壞疽病(病菌Pseudomonas viridiflava、P. mediterranea)萎凋型病害常造成番茄產業的大量損失，且無防治藥劑可推薦，因此本試驗擬調查番茄捲葉病病因之變化，了解抗病品種抗病能力下降的原因，並構築其農桿菌媒介接種系統，定量穩定篩選抗病品種，期能加速抗病品種育成，降低目前因防制粉蝨大量使用農藥所付出的經濟成本。對番茄系統性病原細菌建立鑑定與防治技術，分別針對各病菌篩選適合的藥劑資材，以供產業及農友防治利用。(三)開發農業害蟲寄生蜂之診斷鑑定技術—粉介殼蟲科寄生蜂之形態診斷鑑定本研究將加強臺灣的重要粉介殼蟲之寄生蜂鑑定及分類，增加其生物資源的資料，以供生物防治應用時參考。主要調查對象為膜翅目跳小蜂科 (Encyrtidae) 四突跳小蜂亞科 (Tetracneminae) 的長索跳小蜂族 (Anagyrini) ，針對長索跳小蜂族之屬、種檢索表、形態特性圖、寄主、寄主所在植物及分佈等資料並建立資料庫，以供田間應用時之參考。 五、設施小果番茄非疫生產點建立之評估目前國際間尚無以「設施」(screen house)做為非疫生產點(pest free production sites)之先例，本計畫調查不同設備條件的設施，證明設施可以防堵瓜實蠅( Zeugodacus cucurbitae Coquillett)與東方果實蠅( Bactrocera dorsalis Hendel)入侵，建立以設施做為小果番茄( Lycopersicon esculentum Mill.)非疫生產 點之示範園區，目的為提出設施小果番茄非疫生產點之設備條件。 六、運用智能化排程分析高通量小分子RNA比對重要番茄與番椒種子病原技術建立 本計畫目的是收集與分析小分子RNA序列數據，以建立植物病原偵測的方法。協助國內種子種苗業者檢定重要材料病原，建立健康母本種原庫。本技術可以應用作物新品種材料申請。採樣人員可至輸出國作疫情材料調查。邊境把關人員也可以針對集中輸入樣品，進行病害相分析，確認病害相後，進行去病原處理後，再以RT-PCR等二階檢定技術進行抽樣複檢。本技術預期可加強疫情分析量能，也加速引入育種材料時程。於2020年我們已經收集2種番茄種子與3種植株進行高通量小分子RNA檢定試驗。我們分析發現台灣未報導之番茄病毒。也在輸出入番茄種子檢定出檢疫病原。本年度110年將應用此技術於台灣出口番椒種子進行種傳病原檢定。Investigation and development of technology of diagnosis and identification of pests and pathogens and quarantine treatment for imported and exported plant quarantine</strong><strong>1. Integrated system for identification and analysis of quarantine pests intercepted by border inspection</strong> This expert system is mainly inviting domestic experts to identify and analyze plant pathogens, insects, mites and weeds, etc., that intercepted from import or export agricultural commodities during quarantine inspection and provide the results to the competent authority for adjudication basis. The work aims and expected outcomes of this year's project include 1) Establish a hierarchical responsibility system to identify quarantine pests of imported and exported agricultural commodities to strengthen the domestic pest identification system; 2) Organize training courses on identification techniques of important quarantine pests (mainly mites and aphids) to improve the quarantine personnel's sample handling capacity and species identification; 3) The institutionalization of the handling process and identification plan of the emergency quarantine pest samples can make the overall quarantine identification system more complete and improve quarantine pest identification efficiency. This system will help complete the domestic quarantine and identification system and be consistent with the international system. This project's quarantine objects include plant diseases such as fungi, bacteria, viruses, nematodes, etc., and harmful insects such as hemipteran (aphids, psyllids, and leafhoppers), hymenopteran, lepidopteran pests, etc., and mites.<strong>2. Establishment of diagnostic technique, epidemic information and education training systems of fruit flies (Diptera: Tephritidae) with quarantine importance</strong> This project aims to establish a more concise and accurate fruit fly information and references database, including: 1) Revising fruit fly species checklist in Taiwan. 2) Collecting international fruit fly epidemic data. 3 ) Strengthening fruit fly diagnosis ability and providing pest species information. Confirming fruit fly species of quarantine importance is usually an important issue in agricultural trade negotiations. To prevent unnecessary difficulty during quarantine inspection and trade negotiations, setting up the species diagnostic atlas and species, distribution, and host information will be useful, not only for the quarantine and trade negotiation purpose but also for educational or agricultural perspectives.<strong>3. Molecular identification on thysanopteran, lepidopteran, dipteran, and hemipteran insect pests based on DNA barcodes</strong> Agricultural pests are commonly feeding on crops, which would damage agricultural production. This study would establish and apply DNA barcodes to identify the important agricultural pests in Taiwan and the quarantine pests on importin plant products amongst Thysanoptera, Lepidoptera, Diptera, and Hemiptera. The acquired DNA sequences would be applied as well to design the specific-primers to identify the quarantine pests. Moreover, those important pests' specific DNA probes would be applied and exploited to develop a biochip identification system acquiring efficiency and massive pest identification in the future.<strong>4. Cold quarantine treatment of small-size tomato fruit</strong> The small-size tomato was introduced many years ago. In the cooperation between domestic seed producers and the cultured farmers, the small-size tomato fruit is produced with good quality. In addition to domestic consumption, it can also be exported. However, Taiwan is an epidemic area of fruit flies such as oriental fruit flies, melon flies, and pumpkin fruit flies. Therefore, exporting the tomato fruit to non-epidemic areas requires a quarantine treatment to disinfect the possible fruit flies. The study herein is aimed at the low-temperature treatment to disinfest the three species of fruit flies possibly infected with tomato fruit. And then establish the quarantine treatment for an optimal method to export the tomato fruits.<strong>5. Development irradiation processing technology applied to phytosanitary treatment for export guava fresh fruits</strong> This project aims (1) to use the existing domestic radiation treatment facilities to carry out the experiments of irradiation quarantine treatment of fresh guava (<em>Psidium guajana</em> L) fruit and draft a standard procedure for exported guava quarantine treatments, providing the BAPHIQ as a reference. (2) to continually collect and sort out the established quarantine procedures and regulations to import irradiation-treated fresh fruits in different countries. (3) to explore using molecular biology techniques to examine the biological changes after radiation treatment to confirm whether the fruit is irradiated. Finally, this project is expected to establish the irradiation technology of quarantine treatment of exported guava to reduce fruit damage, expand export opportunities, and increase farmers' benefits. ●<strong>Risk assessment and control measures of important quarantine and emerging pests</strong>
1.International outbreak analysis and development of detection methods for important quarantine viruses In order to maintain the accuracy of our quarantine pest list, we will continue investigating and correcting the classification and scientific name of remaining 169 listed viruses and also add new important quarantine viruses to the pest list. The revised information and collected documents derived from a total of 475 viruses in the pest list will be thoroughly reviewed and reorganized again. Additionally, in recent years, many countries have paid attention to a new seed-borne virus,<em> Tomato mottle mosaic virus</em> (ToMMV), that can infect tomatoes and peppers. We will collect the literatures and sequence information of ToMMV. ToMMV-specific primers will be designed based on viral sequence data. The RT-PCR detection method of ToMMV will be developed accordingly. Finally, the complete revision of quarantine virus list and the detection method for ToMMV will be provided to the authorities of plant inspection and quarantine. 2.Development identification and assessment method of latent quarantine risk nematodes Due to the increasing detection of important plant parasitic nematodes in the inspected importing planting materials and in the planting materials attached to the importing plants, and the need of preventing the entrance of high risk quarantine nematode pests, this project aims to develop suitable quarantine strategy for nematodes in the <em>Aphelenchoides </em>genus. Specifically, this project will analyze the quarantine risks and available detection methods based on the biological, pathological and molecular information of the nematodes, and identify nematode species with latent quarantine risks. This project will utilize the information collect from the scientific publications and from the nematode sample extracted from the import agriculture cargos, and evaluate the feasibility and the efficacy of applying the available detection at the quarantine inspection sites. In addition, to increase the numbers of personals with professional nematode identification expertise, this project also aims to develop a nematologist training course to suit long-term quarantine inspection purposes.3.Assessment of the invasion risk and identification of the emerging lepidopterous pests in the surrounding Asian countriesFall armyworm (Spodoptera frugiperda, FAW) is known as one of the most severe and rapidly increasing risks for more than 370 crops in the world. Due to the fact that FAW suddenly invaded Taiwan in June, 2019, establishemnt of the following knowledge bases gain priority for immediate and future planning for management and control of this pest. We will first simulate the potential geographical expansion and spread of FAW in Taiwan by employing ecologica niche modelling (ENM) and predicted species distribution model (PSD). We then will clarify the identification and diagnosis of FAW and other noctuoid species associated with the crops in Tawian by using morphological characters, including those of adult, larval and pupal stages. We will also collect samples of FAW from the surruonding countried being invaded by this insect in order for better comparison in morphology as well as for future uses in other studies, such as population genetics, genetic basis of resistance, and eludicating the geographical source(s) of the invading population. We wish the results of this project can provide information for future IPM, train people for fast identification, publish scientific reports and offer identificaiton services for govenment agency and farmers. ●<strong>The Study on international surveillance of plant pests epidemic information analysis and tools development.</strong>1.Function-Expansion and Maintenance of Search System of “Approved Plant List for Importation and its Quarantine Requirements” and “Plant Quarantine Pests Database” The phytosanitary regulations are important reference materials for the industry when importing agricultural products from abroad. This study is based on the "Plant Quarantine Pests Database" which was built in 2019, and three tasks are planned: (1) maintain the basic information of the "Plant Quarantine Pests Database".(Including scientific name and classification status); (2) new features of risk assessment profile generation for pests ; (3) new design of "Plant Epidemic Information", which have the functions of automatic knowledge acquisition mechanism of scientific and technological literature. At the same time, the previous study has been completed in accordance with the "Republic of China Imported Plants or Plant Products Quarantine Regulations" amendment and added 768,269 items of provisional data of "Reverse List to Front List"; Continue to maintain a total of 54,353 items in the "Conditional Imported Plant List" database and 35,956 items in the "Prohibited Imported Plant List" database. In order to improve the query accuracy of the public and business colleagues when using "Approved Imported Plant List and Quarantine Condition Query System", the front-end optimization of the search interface of "Public Inquiry" and "Quarantine Officer Inquiry" has completed the diversion and testing of different inquiry pages for user inquiry scenarios. Also, the system's query mechanism optimization, database content and back-end management functions are fully adjusted. Therefore, the project continues to maintain and expand the functions of the "Approved Imported Plant List and Quarantine Condition Inquiry System" and the "Plant Quarantine Pests Database.”At the same time, the previous study has been completed in accordance with the "Republic of China Imported Plants or Plant Products Quarantine Regulations" amendment and added 768,269 items of provisional data of "Reverse List to Front List"; Continue to maintain a total of 54,353 items in the "Conditional Imported Plant List" database and 35,956 items in the "Prohibited Imported Plant List" database. In order to improve the query accuracy of the public and business colleagues when using "Approved Imported Plant List and Quarantine Condition Query System", the front-end optimization of the search interface of "Public Inquiry" and "Quarantine Officer Inquiry" has completed the diversion and testing of different inquiry pages for user inquiry scenarios. Also, the system's query mechanism optimization, database content and back-end management functions are fully adjusted.This year, the epidemic data of orchid diseases and pests will also be added to enrich the database content.2.Information collection and development of control techniques for orchid diseases and pests Taiwan's mainly export orchids are <em>Phalaenopsis</em>, <em>Cymbidium</em> and <em>Cattleya</em> and so on. Those orchids have export pressures from international competition, and enhancement of yield and quality is one of the feasible ways to increase competitive advantage. However, during the cultivation of orchids, the occurrence of pests and diseases is one of the main reasons for the decline of yield and quality. In the past, the development of a viable pest management strategy was following specific single disease or pest. But, practitioners may misunderstand or don't know about the occurrence of pest and disease ecology when they faced with different diseases and pests. This not only failed to remedy the problem but also delayed the best time to prevent pests. In addition, the occurrence of pests and diseases, especially the presence of pests, often leads to export barriers and poor quality of delivery at the time of export quarantine. Focusing on the perspective of clean seedling system and pest management in the greenhouse, this study will aim at development an integrated pest management in common pests and diseases of <em>Phalaenopsis</em>, <em>Cymbidium</em> and <em>Cattleya</em>. ●<strong>Development and application of diagnosis and identification of crop diseases and pests</strong>
1.Chrysomelinae (Coleoptera: Chrysomelidae) in the agroecosystem: taxnomy, host plants, and population dynamics under climate change Chrysomelidae is one of the most diverse families of Coleoptera. More than six hundred species have been described or recorded from Taiwan. However, most of taxonomic studies on chrysomelids were restricted to short descriptions of new species or announcement of new records. The collecting sites of the studied specimens were always scattered and the number of the studied specimens were fewer as expected. This research is dedicated to the modern taxonomic studies on Chrysomelidae of Taiwan. The species belonging to the subfamily Chrysomelinae are revised comprehensively. Specimens will be collected intensively and extensively. Diagnostic characters will be searched throughout all members of the genus. Adult appearance, host plants, and the geographic distribution of each species will be major components of this research. The type specimen of the known species will be reexamined. A sound classification of Chrysomelidae in Taiwan is expected to be established.2.Development of identification and control techniques for the tomato systemic diseases Tomato crop is one of the important vegetable in Taiwan. Systemic diseases are often caused by viruses and bacteria, causing severe loss. At present, the main virus diseases are tomato leaf curl and yellow leaf curl caused by geminivirus infection. The main control measures are resistant of tomato varieties to vector insect whitefly. .However, the investigation found that the commonly mixed infection or genetic recombination of geminiviruses in tomato, which makes the virus with high mutation rate and the disease resistance of tomato varieties with disease resistance gene is decreasing year by year.Because whitefly has adapted to Taiwan's natural environment and has a strong omnivorous nature, in order to control the disease with pesticides, it must be applied intensively. The breeding of resistant varieties is still the most effective and environmentally friendly method. Currently, for the selection of resistant varieties the plants are still placed in the net room and infected with whitefly vector. The screening results differed according to the whitefly population and the transmission efficiency, which increased the experimental variabilities. In addition, many bacterial diseases of tomato could be found in the field. The bacterial pathogen included <em>Ralstonia solanacearum</em>, <em>Pseudomonas viridiflava</em>, and <em>P. mediterranea</em> could result in wilt symptom on tomato and caused large losses. There are few control agents recommended.Therefore, this study intends to investigate the symptoms change of tomato leaf curl disease, understand the reasons for the decline in disease resistance of disease-resistant varieties, and build its Agrobacterium-mediated inoculation system to quantitatively and stably screen disease-resistant varieties. For tomato systemic pathogenic, this study establishes the identification and control technology of bacteria, and screen control materials for each pathogen for industry and farmers to control bacterial diseases. 3.Development of diagnostic and identification techniques for parasitoids of agricultural pests- parasitoids of mealybugs This project intends to establish the toxonomic system of Anagyrinian parasitoids reared from mealybugs in Taiwain. All of the Anagyrini of Taiwan, belong to the subfamily of Tetracneminae (Hymenopera : Encyrtidae). All specimens described will be deposited in the Taiwan Agricultural Research Institute (TARI), Taichung, Taiwan. Furthermore, keys to genera and species of Anagyrini are provided along with illustrated figures and bionomic notes such as data of host, host associated plants and their distribution in Taiwan. The research will set up the data of parasitic wasps in biological resources and enhance the biocontrol efficiency for important mealybugs in Taiwan. ●<strong>The assessment of the establishment of pest free production sites for cherry tomatoes in screen houses</strong> At present, no example is available in the world to use "screen houses" as a pest free production site. The research evaluated the feasibility of using screen houses to approve for the complete prevention of melon fly ( Zeugodacus cucurbitae Coquillett) and fruit fly ( Bactrocera dorsalis Mill.). The final goal is to propose an ideal design of screen houses for a pest free production site of cherry tomatoes, and to establish demonstration farms. ●<strong>The establishment of high through-put sequencing workflow to detect pathogens of tomato and Capsicum seeds</strong> The purpose of this project is to collect and analyze small molecule RNA sequence data to establish a method for plant pathogen detection. Assist domestic seed and seedling operators to verify the pathogens of important materials and establish a source database of healthy species. This technology can be applied to applications for new crop varieties. Samplers can go to the exporting country to investigate the epidemic materials. Border guards can also conduct disease phase analysis for centralized input samples, and after confirming the disease phase, conduct de-pathogen treatment, and then conduct sampling re-inspection using second-level verification techniques such as RT-PCR. This technology is expected to enhance the capacity of epidemic analysis and speed up the introduction of breeding materials. In 2020, we have collected 2 lots tomato seeds and 3 diseased plants for high-throughput small molecule RNA assays. Our analysis revealed unreported tomato viruses in Taiwan. The quarantine pathogen was also detected in the import and export of tomato seeds. In 2021, we will apply and improve the analysis process to verify the seed-borne pathogens of the pepper. |
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