鑑定稠李鏈黴菌PMS-702 作為生物農藥的菌種

A Streptomyces PMS-702 strain was isolated from farmland soil collected from Taiwan. Antimicrobial activities of PMS-702 strain against phytopathogenic fungi and bacteria showed differential inhibitory effects on the growth of fungal pathogens, but no effect for Sclerotium rolfsii and nine plant pathogenic bacteria. According to the results of morphological, physiological and biochemistry characteristics, and 16S rRNA gene sequence, the PMS-702 strain was identified as Streptomyces padanus. Biomass of S. padanus PMS-702 strain varied among different nutrient media for growing. Soybean Meal-Glucose Broth (SMG) medium was the best for enhancing the growth of S. padanus strain PMS-702 and its crude culture filtrate diluted to 10 fold completely suppressed spore germination of Fusarium oxysporum f. sp. cubense, which was used as an indicator for assessing antimicrobial activities. In bioassay experiment, the culture filtrates of S. padanus from SMG cultures fermented at 30 degrees Centigrade for 4 days could control effectively a broad range of plant diseases including lettuce brown spot, mango anthracnose, Chinese cabbage anthracnose, peach fruit rot, and orange green mold. These results suggested that S. padanus strain PMS-702 is potential to be used as a biological control agent to suppress fungal crop diseases. 放線菌PMS-702 菌株係由台灣農田中分離所獲得，其對於各種供試植物病原真菌具有不同程度的拮抗能力，對於 白絹病菌 (Sclerotium rolfsii) 與9 株植物病原細菌則均不具有拮抗的效果。分析PMS-702 菌株之形態、生理、生化特 徵及16S rRNA 基因序列後，確定PMS-702 菌株為稠李鏈黴菌 (Streptomyces padanus)。PMS-702 菌株在不同營養配方培養具有差異化，以黃豆粉葡萄糖培養基 (SMG) 培養PMS-702 具有最佳的生質量，其10 倍稀釋培養粗濾液具有抑制香蕉黃葉病菌 (Fusarium oxysporum f. sp. cubense) 孢子發芽的效果。PMS-702 菌株在SMG 培養基於30oC 培養4 天後，利用生物檢定分析法測試其對於數種植物病害的影響，結果發現PMS-702 菌株培養濾液對於萵苣褐斑病、芒果炭疽病、白菜炭疽病、甜桃果實疫病及椪柑果實綠黴病均具有防治功效，這些結果顯示稠李鏈黴菌PMS-702 具有研發成為生物製劑，用於防治作物真菌性病害的潛力

Integrated Pest Management for Important Plants

由於台灣地處亞熱帶，氣候高溫多濕，田間常發生病蟲害造成農民損失，因此如何整合一套有效病蟲害管理辦法是重要課題。本計畫的目的在於統合不同植物病蟲害領域專長之專家，參與台灣地區重要疫病蟲害之管理。計畫工作項目包括，田間重要作物病蟲害種類之調查、主要病原微生物與有害昆蟲之鑑定、有益微生物之篩選與應用、有益植物萃取液之篩選與應用、田間主要病原微生物與有害昆蟲之抗藥性監測及致病基因探討等。本計畫亦將定期對農民進行教育訓練，並透過政府與各地方之縣市政府舉辦重要疫病蟲害整合管理觀摩會。Taiwan locate in sub-tropical area, and climate is high temperature and humidity. Therefore, pests and diseases often cause farmers' ecomonic losses. Thus, developing an effective integrated pest management approach is an important issue. The objective of this project is integration of different expertise in the plant pathologist and entomologist to participate in an important disease management of pests and diseases in Taiwan. The works is including the survying of important crop pests and diseases in field, identification of main pathogenic microorganisms and harmful insects, screening and application of beneficial microorganismsf, screening and application of benificial plant extracts, monitor of resistant pathogens and insects in field, investigating the pathogenic genes. This project will also be regular seting up training program for farmers, and the observing will be integrated the management of plant diseases and insect pests by the goverment and the local governments

重要植物疫病蟲害整合性管理

Taiwan locate in sub-tropical area, and climate is high temperature and humidity. Therefore, pests and diseases often cause farmers' ecomonic losses. Thus, developing an effective integrated pest management approach is an important issue. The objective of this project is integration of different expertise in the plant pathologist and entomologist to participate in an important disease management of pests and diseases in Taiwan. The works is including the survying of important crop pests and diseases in field, identification of main pathogenic microorganisms and harmful insects, screening and application of beneficial microorganismsf, screening and application of benificial plant extracts, monitor of resistant pathogens and insects in field, investigating the pathogenic genes. This project will also be regular seting up training program for farmers, and the observing will be integrated the management of plant diseases and insect pests by the goverment and the local governments.由於台灣地處亞熱帶，氣候高溫多濕，田間常發生病蟲害造成農民損失，因此如何整合一套有效病蟲害管理辦法是重要課題。本計畫的目的在於統合不同植物病蟲害領域專長之專家，參與台灣地區重要疫病蟲害之管理。計畫工作項目包括，田間重要作物病蟲害種類之調查、主要病原微生物與有害昆蟲之鑑定、有益微生物之篩選與應用、有益植物萃取液之篩選與應用、田間主要病原微生物與有害昆蟲之抗藥性監測及致病基因探討等。本計畫亦將定期對農民進行教育訓練，並透過政府與各地方之縣市政府舉辦重要疫病蟲害整合管理觀摩會

Survey, Exploitation and Application of Biocontrol

本計畫的目的在於： 1. 植物傷口癒合防護劑之研發與應用-篩選有益傷口癒合防護之微生物與拮抗菌,開發評估平台並合成傷口癒合防護劑。 2. 釋放捕食性天敵防治蘇鐵白輪盾介殼蟲及其效果評估-了解生物天敵雙色出尾蟲捕食能力，做為釋放天敵數量之依據。大量飼養蘇鐵白輪盾介殼蟲之生物天敵雙色出尾蟲以建立蘇鐵白輪盾介殼蟲生物天敵族群，適合野外蘇鐵之生物防治模式，減少未來每年為防治蘇鐵白輪盾介殼蟲為害所付出之成本以完成蘇鐵白輪盾介殼蟲之綜合防治模式，使全國各地之蘇鐵白輪盾介殼蟲疫情得以控制，避免台東蘇鐵林滅絕。 3. 芒果單爪葉蟎(Oligonychus mangiferus)為芒果重要的害蟲之ㄧ，同時危害荔枝、葡萄、梨與桃等多種作物。卵形捕植蟎(Amblyseius ovalis)於熱帶果樹分布廣泛，且可以捕食大量單爪葉蟎，值得評估其生物防治利用價值。本研究擬研發與現行蟲害管理系統相容之監測與生物防治技術，供擬定芒果防疫對策之參考。 建立循序與二項分布取樣法可應用於田間觀測，且具省時、省力、準確估計芒果單爪葉蟎族群密度等特性，利於監測芒果單爪葉蟎族群終年發生頻度。 利用捕植蟎施用密度與數量反應與捕植蟎施用時間與葉蟎族群壓制力，可有效擬訂施用生物防治的適當時機與施用密度。捕植蟎擴散與控制能力研究結果，有助捕植蟎之田間利用與施用方法之改進，進而提供今後生物防治芒果單爪葉蟎之技術開發，落實非農藥防治及生物技術之利用，提高農友經濟收益，促進產品外銷及提昇我國農產品競爭力。 4. 豆莢螟幼蟲喜暱食於嫩芽、花器及豆莢，藥液常難觸及，容易增加用藥次數及用量；而豆類蔬菜生長快速，採收期長，用藥不可不慎，為提升豆類蔬菜之品質，減少農藥用藥用量及殘留量，擬綜合運用微生物、生物防治及性費洛蒙等防治技術，共同防治豆莢螟。本年度計畫之工作重點為： (1) 搭建可拆卸式網室，種植長豇豆，釋放豆莢螟及寄生蠅，評估寄生蜂及寄生蠅於網室長豇豆之寄生防治效果，此試驗於春、秋兩季節進行。 (2) 搭建可拆卸式網室，評估性費洛蒙於長豇豆試驗田，對田間豆莢螟之干擾交尾與產卵效果。 (3) 單獨使用豆莢螟核多角體病毒(BA)、或與蘇力菌、苦楝油、寄生蜂混用，於長豇豆及鵲豆試驗田，評估對豆莢螟之綜合防治效果，此試驗亦於春、秋兩季進行試驗。 5. 目前在植物病害的防治上多依賴化學農藥，長期大量的使用不僅加劇環境污染，更直接危害人類健康。如何以非農藥防治方法，降低對化學農藥之使用，提高我國重要農產之競爭力，是當今病害防治工作的重要方向。本計畫將收集並評估可以防治植物寄生性線蟲及抑制芒果、木瓜、蓮霧、草莓之重要病原真菌的植物種類；在殺線蟲劑的測試使用先前初步試驗有效果之天人菊，以不同施用量與型態(生鮮植體碎片、乾燥碎片)處理感染根瘤線蟲的蕹菜，測試其防治效果；並觀察對於蕹菜、番茄、絲瓜等作物的生長是否有影響。且蒐集其它有毒或無毒的觀賞植物，使用水溶性粗萃取液處理南方根瘤線蟲(Meloidogyne incognita)，亦處理危害芒果、木瓜、蓮霧、草莓之病原微生物；評估不同植物抽出物與化學藥劑防治病害之效果差異。作為開發防治之生物農藥，減少劇毒性殺線蟲劑之使用，降低線蟲或病原真菌產生抗藥性速率及生態環境之破壞。 6. 臘狀芽孢桿菌C1菌株分離自百合根圈，已知其施用於百合根圈可增加植株對灰黴病之抵抗性，本計畫擬應用所分離之臘狀芽孢桿菌C1菌株進行其在他種作物病害之防治試驗，瞭解其應用潛力；針對玉米葉枯病進行試驗，測試其施用方式，進行田間試驗及瞭解其使農藥減量之效果，以評估C1菌株與殺菌劑共同使用之可行性。 7. 利用拮抗細菌(Bacillus subtilis LB5)之液態培養稀釋液、木黴菌(Trichoderma sp.)固態發酵液、蘇力菌、印楝素、印楝油及無患子油從蓮霧生長期中，每隔10-14天噴施地上部及地下部落葉，配合中果期之套袋，期以獲得蓮霧病蟲害之非農藥防治效果之技術。調查蓮霧生長期之葉部枝條病菌害蟲及土壤枯枝落葉病菌及空中病菌密度，以了解非農藥處理之田間病菌及害蟲變動情形。 8. 由於番石榴粉介殼蟲在台灣危害寄主作物甚廣，超過250種。包括番石榴、柑桔、玉米和葡萄等重要作物。因此番石榴粉介殼蟲的防治相當重要。為了要避免因使用農藥來防治，所產生的不良副作用，改選用無毒性的性費洛蒙來誘捕是一有效且可行的防治方法。最近番石榴粉介殼蟲之性費洛蒙已被分離、鑑定為(E)-2-isopropyl-5-methyl-2,4-hexadienyl acetate。因此本計畫乃要研發一新簡便的合成方法來合成此費洛蒙並提供作田間誘捕和監測之用，以有效防治此蟲。The purposes of the study are to 1. Development of bioprotective agent for healing plant wounds. 2. Releases and evaluation of the effectiveness of a predtor, Cybocephalus binotatus, in controlling the cycad scale, Aulacaspis yasumatsui - In order to establish a natural enemy population as the biological control on the field, the mass propagation of C. japonicus will be conducted. In addition, the capture ability of C. japonicus to A. yasumatsui will be studied as the reference for mass release. The results will provide a completed pest manage model and reduce the total cost of controlling A. yasumatsui, benefiting the whole nation and avoiding the extinction of Cycad forest in Taitung. 3. 4. The legume pod borer (LPB), Maruca vitrata (F.) (syn. M. testulalis) (Lepidoptera: Pyralidae) is an important pest affecting vegetable and grain legumes in Taiwan. As the LPB larvae feed on tender shoots, flowers and pods by concealing inside, it is very difficult to control them adopting a single approach. At present, farmers rely exclusively on the application of chemical insecticides to combat LPB, especially on vegetable legumes. The over reliance on chemical insecticides has posed insecticide residues in the fresh vegetables because they are harvested in short intervals of two or three days. Therefore, an integrated approach, based on bio-pesticides, natural enemies and sex pheromones, is proposed. Major activities and implementation approaches include (i) testing the individual and combined effectiveness of parasitoids Nemorilla maculosa and Apanteles taragamae against LPB larvae in yard-long bean field during spring and autumn season, (ii) testing the mating and oviposition disruption of the LPB sex pheromones in yard-long bean field, (iii) testing the effectiveness of MaviMNPV formulations alone and in combination with Bt, neem and parasitoids, and (iv) testing the IPM strategy with parasitoids, MaviMNPV, neem and Bt in field experiments with yard-long bean and lablab bean. Thus, this proposal aimed to exploit the bio-control based IPM strategy to combat the LPB in vegetable legumes, which would not only reduce the insecticide use but also enhance the efficiency of natural enemies in field conditions. 5. Pesticides are the most relied on control tactic, but they had great draw backs on environmental safety and human health. One of the goals of disease management is to integrate non-chemical control tactics to lower pesticides usage and increase the revenue of farm products. Natural herb would be collected and tested for the nemacidal and fungicidal efficacy. Gaillardia plucella had good nemacidal efficacy in the preliminary tests and its different formulation as well as quantity of application would be further tested in this project on root-knot infected water spinach. The Gaillardia plucella formula would also be tested for its effect on tomato and gourd growth. Natural herbs are collected and their extraction would be twsted against fungal pathogens on mango, papaya, wax apple and strawberry and compared with the efficacy of conventional fungicides. The purpose of the research is to lower the amount of pesticides used in the future and the risk of pathoges to develop pesiticide-resistance. 6.7. Bacillus cereus C1 was isolated from the rhizosphere of lily plants. When the bacterial suspension was applied to the plant rhizosphere, the incidence of Botrytis leaf blight was decreased. The goal of this project is to test the efficacy of B. cereus C1 on the control of other plant diseases. Corn leaf blight is the disease target. The application method, field trials, the reduction efficiency on fungicide usage will be examined to evaluate the efficacy of C1 application and the feasibility of application combination with fungicide. 8. For developing the non-pesticide control techniques for management of plant diseases and pests of waxapple, the antagonistic bacterium(Bacillus subtilis LB5) and fungi (Trichoderma sp.), commercial biopesticide Bt, azadirachtin are sprayed at 10-14 days interval in waxapple orchard during growing stage , and then integrated with fruit bagging after fruitlet stage. The fungal pathogens and insect pests are investigated preharvest and postharvest to evaluate the control efficiency of the non-pesticide method. 9. Due to the passionvine mealybug, Planococcus minor (Maskell), is a serious pest of more than 250 host plants, including major agricultural crops as diverse as guava, citrus, corn, grapes and tree fruits in Taiwan, the effective control of this pest is very important and has attracted considerable attention. To avoid the side effects occurring by using pesticides, the nontoxic sex pheromone of the passionvine mealybug is chosen instead. Recently, the sex pheromone of this insect has been identified as (E)-2-isopropyl-5-methyl-2,4-hexadienyl acetate. The purpose of this project is to develop a new convenient method to synthesize this sex pheromone and provide it for the mass trapping and monitoring of this pest. Accordingly, the effective control of this pest will be successfully achieved in the near future

Survey, Exploitation and Application of Biocontrol Agent Resources for Management of Plant Pests

本計畫的目的在於： 1. 植物傷口癒合防護劑之研發與應用-篩選有益傷口癒合防護之微生物與拮抗菌,開發評估平台並合成傷口癒合防護劑。 2. 利用枯草桿菌生物製劑防治芒果及木瓜炭疽病可行性之探討-研究目標將以實驗室篩選出之本土優良拮抗枯草桿菌BS-99菌株，進行芒果及木瓜炭疽病的田間防治試驗。 3. 利用有益微生物與熱能防治外銷花卉根部病害之研究-利用熱能及本土性之微生物來減少洋秸梗病害及降低化學藥劑之使用。 4. 利用土壤添加物及拮抗微生物防治長豇豆萎凋病-以拮抗微生物配合土壤添加物，嘗試配合種子粉衣、剪胚軸接種或育苗介質混菌等方法，將拮抗微生物導入長豇豆幼苗，供作田間防治之用。 5. 益生性枯草桿菌在柑橘潰瘍病綜合防治管理之應用-完成預期可建立完成可專利化之「枯草桿菌生物殺菌劑泛用性FNF產製與應用」智財權技術，其推廣應用後除可強化我國農業生產之國際競爭力，並將加速永續農業政策推動之全面落實。 6. 植物病害防治用廣效性微生物製劑之研發與應用-發展螢光假單胞菌製劑來防治細菌性病害。 7. 利用有毒植物萃取液防治作物線蟲病害-評估有毒植物萃取液對不同植物寄生性線蟲之影響與應用於溫室之防治植物寄生性線蟲效果。 8. 利用產氰細菌防治作物線蟲病害之效果評估。 9. 芒果單爪葉蟎生物防治天敵效能評估-捕植蟎捕食芒果單爪葉蟎之功能反應之結果有助捕植蟎之田間利用，進而提供今後生物防治芒果單爪葉蟎之技術開發，落實非農藥防治及生物技術之利用，提高農友經濟收益，促進產品外銷及我國農產品競爭力。 10. 豆莢螟綜合防治技術之研究與開發(亞洲蔬菜中心-施瑞霖) (1) 開發豆莢螟寄生蜂Apanteles taragamae之大量飼養技術，及探討其生物學特性，包括最適寄生之寄主齡期、最適寄主密度、對溫度的敏感度等。 (2) 測試豆莢螟性費洛蒙之效果。 (3) 搭建可拆卸式網室，種植長豇豆，釋放豆莢螟及寄生蠅，並評估寄生蠅於網室長豇豆之寄生效果。 (4) 利用殺蟲效果較佳之豆莢螟核多角體病毒製劑BA與BH，單獨使用或與其他生物性藥劑(如蘇力菌、苦楝油)混用，評估豆莢螟核多角體於田間之防治效果。 (5) 綜合豆莢螟寄生蠅或寄生蜂、豆莢螟核多角體病毒、蘇力菌及性費洛蒙等防治技術，應用於長豇豆試驗田，並評估對豆莢螟之防治效果。 11. 釋放捕食性天敵防治蘇鐵白輪盾介殼蟲及其效果評估-了解生物天敵雙色出尾蟲捕食能力，做為釋放天敵數量之依據。大量飼養蘇鐵白輪盾介殼蟲之生物天敵雙色出尾蟲以建立蘇鐵白輪盾介殼蟲生物天敵族群，適合野外蘇鐵之生物防治模式，減少未來每年為防治蘇鐵白輪盾介殼蟲為害所付出之成本以完成蘇鐵白輪盾介殼蟲之綜合防治模式，使全國各地之蘇鐵白輪盾介殼蟲疫情得以控制，避免台東蘇鐵林滅絕。The purposes of the study are to 1.Development of bioprotective agent for healing plant wounds. 2.A study on microbial formulation of Bacillus subtilis for biological control of mango and papaya anthracnose diseases by Colletotrichum gloeosporioides.- The objectives of this study are to evaluate the disease control potential of Bacillus subtilis BS-99 strain against anthracnose disease causing by Colletotrichum gloeosporioides, on mango and papaya under field conditions. 3.Root disease control of export flowers by heat and effective microorganisms- In order to reduce crop diseases and reduction of chemical pesticide, the study in this project will conduct to use the steam with 70℃ and antagonists to control nematodes and root rot disease of export flowers. 4.Studies on soil amendments and antagonists for controlling Fusarium wilt of asparagus bean- Seeds and seedlings of asparagus bean will be treated with the potential biocontrol agents and planted into the pathogen-infested soils in the greenhouse and in the field. Based on the results obtained, we try to formulate a soil amendment with antagonists for controlling Fusarium wilt of asparagus bean in the field. 5. Application of probiotic Bacillus subtilis biofungicide in intergrated management of citrus canker disease- The completion of the project should provide a patentable proprietary technology for producing FNF from available B. subtilis biofungicides useful for sustainable agricultural production. 6.Development and application of broad-spectrum microbial agents for controlling of plant diseases- develop formulations of fluorescent Pseudomonas to control plant bacterial diseases. 7.Application of extracts of poisonous plant for control of nematode disease- Effective ingredients would be tested under greenhouse condition for its potential for future application. 8.Studies on the management of plant-parasitic nematode disease by using of cyanogenic bacteria. 9. 10. Developing sustainable IPM strategy for the control of legume pod borer, Maruca vitrata in vegetable legumes-(1) developing and standardizing reproducible mass culturing techniques for A. taragamae, (2) laboratory and greenhouse studies on the nature and extent of the parasitism by A. taragamae on LPB larvae viz., the most suitable host larval stage, effect of host larval population density on parasitism and effect of temperature, to optimize the conditions for field release, (3) testing the individual and combined effectiveness of N. maculosa and A. taragamae against LPB larvae field conditions, (4) testing the mating disruption and mass trapping potential of the LPB sex pheromones under controlled and open environment conditions using new pheromone formulations, (5) testing the effectiveness of MaviMNPV formulations alone and in combination with Bt and neem and (6) developing and testing the IPM with parasitoids, MaviMNPV, neem, Bt and sex pheromones in field experiments with yard-long bean. Thus, this proposal aimed to exploit the bio-control based IPM strategy to combat the LPB in vegetable legumes, which would not only reduce the insecticide use but also enhance the efficiency of natural enemies in field conditions. 11. Releases and evaluation of the effectiveness of a predtor, Cybocephalus binotatus, in controlling the cycad scale, Aulacaspis yasumatsui- In order to establish a natural enemy population as the biological control on the field, the mass propagation of C. japonicus will be conducted. In addition, the capture ability of C. japonicus to A. yasumatsui will be studied as the reference for mass release. The results will provide a completed pest manage model and reduce the total cost of controlling A. yasumatsui, benefiting the whole nation and avoiding the extinction of Cycad forest in Taitung

加強生物防治資源調查開發利用研究

The purposes of the study are to 1. Development of bioprotective agent for healing plant wounds. 2. Releases and evaluation of the effectiveness of a predtor, Cybocephalus binotatus, in controlling the cycad scale, Aulacaspis yasumatsui - In order to establish a natural enemy population as the biological control on the field, the mass propagation of C. japonicus will be conducted. In addition, the capture ability of C. japonicus to A. yasumatsui will be studied as the reference for mass release. The results will provide a completed pest manage model and reduce the total cost of controlling A. yasumatsui, benefiting the whole nation and avoiding the extinction of Cycad forest in Taitung. 3. 4. The legume pod borer (LPB), Maruca vitrata (F.) (syn. M. testulalis) (Lepidoptera: Pyralidae) is an important pest affecting vegetable and grain legumes in Taiwan. As the LPB larvae feed on tender shoots, flowers and pods by concealing inside, it is very difficult to control them adopting a single approach. At present, farmers rely exclusively on the application of chemical insecticides to combat LPB, especially on vegetable legumes. The over reliance on chemical insecticides has posed insecticide residues in the fresh vegetables because they are harvested in short intervals of two or three days. Therefore, an integrated approach, based on bio-pesticides, natural enemies and sex pheromones, is proposed. Major activities and implementation approaches include (i) testing the individual and combined effectiveness of parasitoids Nemorilla maculosa and Apanteles taragamae against LPB larvae in yard-long bean field during spring and autumn season, (ii) testing the mating and oviposition disruption of the LPB sex pheromones in yard-long bean field, (iii) testing the effectiveness of MaviMNPV formulations alone and in combination with Bt, neem and parasitoids, and (iv) testing the IPM strategy with parasitoids, MaviMNPV, neem and Bt in field experiments with yard-long bean and lablab bean. Thus, this proposal aimed to exploit the bio-control based IPM strategy to combat the LPB in vegetable legumes, which would not only reduce the insecticide use but also enhance the efficiency of natural enemies in field conditions. 5. Pesticides are the most relied on control tactic, but they had great draw backs on environmental safety and human health. One of the goals of disease management is to integrate non-chemical control tactics to lower pesticides usage and increase the revenue of farm products. Natural herb would be collected and tested for the nemacidal and fungicidal efficacy. Gaillardia plucella had good nemacidal efficacy in the preliminary tests and its different formulation as well as quantity of application would be further tested in this project on root-knot infected water spinach. The Gaillardia plucella formula would also be tested for its effect on tomato and gourd growth. Natural herbs are collected and their extraction would be twsted against fungal pathogens on mango, papaya, wax apple and strawberry and compared with the efficacy of conventional fungicides. The purpose of the research is to lower the amount of pesticides used in the future and the risk of pathoges to develop pesiticide-resistance. 6.7. Bacillus cereus C1 was isolated from the rhizosphere of lily plants. When the bacterial suspension was applied to the plant rhizosphere, the incidence of Botrytis leaf blight was decreased. The goal of this project is to test the efficacy of B. cereus C1 on the control of other plant diseases. Corn leaf blight is the disease target. The application method, field trials, the reduction efficiency on fungicide usage will be examined to evaluate the efficacy of C1 application and the feasibility of application combination with fungicide. 8. For developing the non-pesticide control techniques for management of plant diseases and pests of waxapple, the antagonistic bacterium(Bacillus subtilis LB5) and fungi (Trichoderma sp.), commercial biopesticide Bt, azadirachtin are sprayed at 10-14 days interval in waxapple orchard during growing stage , and then integrated with fruit bagging after fruitlet stage. The fungal pathogens and insect pests are investigated preharvest and postharvest to evaluate the control efficiency of the non-pesticide method. 9. Due to the passionvine mealybug, Planococcus minor (Maskell), is a serious pest of more than 250 host plants, including major agricultural crops as diverse as guava, citrus, corn, grapes and tree fruits in Taiwan, the effective control of this pest is very important and has attracted considerable attention. To avoid the side effects occurring by using pesticides, the nontoxic sex pheromone of the passionvine mealybug is chosen instead. Recently, the sex pheromone of this insect has been identified as (E)-2-isopropyl-5-methyl-2,4-hexadienyl acetate. The purpose of this project is to develop a new convenient method to synthesize this sex pheromone and provide it for the mass trapping and monitoring of this pest. Accordingly, the effective control of this pest will be successfully achieved in the near future.本計畫的目的在於： 1. 植物傷口癒合防護劑之研發與應用-篩選有益傷口癒合防護之微生物與拮抗菌,開發評估平台並合成傷口癒合防護劑。 2. 釋放捕食性天敵防治蘇鐵白輪盾介殼蟲及其效果評估-了解生物天敵雙色出尾蟲捕食能力，做為釋放天敵數量之依據。大量飼養蘇鐵白輪盾介殼蟲之生物天敵雙色出尾蟲以建立蘇鐵白輪盾介殼蟲生物天敵族群，適合野外蘇鐵之生物防治模式，減少未來每年為防治蘇鐵白輪盾介殼蟲為害所付出之成本以完成蘇鐵白輪盾介殼蟲之綜合防治模式，使全國各地之蘇鐵白輪盾介殼蟲疫情得以控制，避免台東蘇鐵林滅絕。 3. 芒果單爪葉蟎(Oligonychus mangiferus)為芒果重要的害蟲之ㄧ，同時危害荔枝、葡萄、梨與桃等多種作物。卵形捕植蟎(Amblyseius ovalis)於熱帶果樹分布廣泛，且可以捕食大量單爪葉蟎，值得評估其生物防治利用價值。本研究擬研發與現行蟲害管理系統相容之監測與生物防治技術，供擬定芒果防疫對策之參考。 建立循序與二項分布取樣法可應用於田間觀測，且具省時、省力、準確估計芒果單爪葉蟎族群密度等特性，利於監測芒果單爪葉蟎族群終年發生頻度。 利用捕植蟎施用密度與數量反應與捕植蟎施用時間與葉蟎族群壓制力，可有效擬訂施用生物防治的適當時機與施用密度。捕植蟎擴散與控制能力研究結果，有助捕植蟎之田間利用與施用方法之改進，進而提供今後生物防治芒果單爪葉蟎之技術開發，落實非農藥防治及生物技術之利用，提高農友經濟收益，促進產品外銷及提昇我國農產品競爭力。 4. 豆莢螟幼蟲喜暱食於嫩芽、花器及豆莢，藥液常難觸及，容易增加用藥次數及用量；而豆類蔬菜生長快速，採收期長，用藥不可不慎，為提升豆類蔬菜之品質，減少農藥用藥用量及殘留量，擬綜合運用微生物、生物防治及性費洛蒙等防治技術，共同防治豆莢螟。本年度計畫之工作重點為： (1) 搭建可拆卸式網室，種植長豇豆，釋放豆莢螟及寄生蠅，評估寄生蜂及寄生蠅於網室長豇豆之寄生防治效果，此試驗於春、秋兩季節進行。 (2) 搭建可拆卸式網室，評估性費洛蒙於長豇豆試驗田，對田間豆莢螟之干擾交尾與產卵效果。 (3) 單獨使用豆莢螟核多角體病毒(BA)、或與蘇力菌、苦楝油、寄生蜂混用，於長豇豆及鵲豆試驗田，評估對豆莢螟之綜合防治效果，此試驗亦於春、秋兩季進行試驗。 5. 目前在植物病害的防治上多依賴化學農藥，長期大量的使用不僅加劇環境污染，更直接危害人類健康。如何以非農藥防治方法，降低對化學農藥之使用，提高我國重要農產之競爭力，是當今病害防治工作的重要方向。本計畫將收集並評估可以防治植物寄生性線蟲及抑制芒果、木瓜、蓮霧、草莓之重要病原真菌的植物種類；在殺線蟲劑的測試使用先前初步試驗有效果之天人菊，以不同施用量與型態(生鮮植體碎片、乾燥碎片)處理感染根瘤線蟲的蕹菜，測試其防治效果；並觀察對於蕹菜、番茄、絲瓜等作物的生長是否有影響。且蒐集其它有毒或無毒的觀賞植物，使用水溶性粗萃取液處理南方根瘤線蟲(Meloidogyne incognita)，亦處理危害芒果、木瓜、蓮霧、草莓之病原微生物；評估不同植物抽出物與化學藥劑防治病害之效果差異。作為開發防治之生物農藥，減少劇毒性殺線蟲劑之使用，降低線蟲或病原真菌產生抗藥性速率及生態環境之破壞。 6. 臘狀芽孢桿菌C1菌株分離自百合根圈，已知其施用於百合根圈可增加植株對灰黴病之抵抗性，本計畫擬應用所分離之臘狀芽孢桿菌C1菌株進行其在他種作物病害之防治試驗，瞭解其應用潛力；針對玉米葉枯病進行試驗，測試其施用方式，進行田間試驗及瞭解其使農藥減量之效果，以評估C1菌株與殺菌劑共同使用之可行性。 7. 利用拮抗細菌(Bacillus subtilis LB5)之液態培養稀釋液、木黴菌(Trichoderma sp.)固態發酵液、蘇力菌、印楝素、印楝油及無患子油從蓮霧生長期中，每隔10-14天噴施地上部及地下部落葉，配合中果期之套袋，期以獲得蓮霧病蟲害之非農藥防治效果之技術。調查蓮霧生長期之葉部枝條病菌害蟲及土壤枯枝落葉病菌及空中病菌密度，以了解非農藥處理之田間病菌及害蟲變動情形。 8. 由於番石榴粉介殼蟲在台灣危害寄主作物甚廣，超過250種。包括番石榴、柑桔、玉米和葡萄等重要作物。因此番石榴粉介殼蟲的防治相當重要。為了要避免因使用農藥來防治，所產生的不良副作用，改選用無毒性的性費洛蒙來誘捕是一有效且可行的防治方法。最近番石榴粉介殼蟲之性費洛蒙已被分離、鑑定為(E)-2-isopropyl-5-methyl-2,4-hexadienyl acetate。因此本計畫乃要研發一新簡便的合成方法來合成此費洛蒙並提供作田間誘捕和監測之用，以有效防治此蟲

加強生物防治資源調查、開發利用研究

The purposes of the study are to 1.Development of bioprotective agent for healing plant wounds. 2.A study on microbial formulation of Bacillus subtilis for biological control of mango and papaya anthracnose diseases by Colletotrichum gloeosporioides.- The objectives of this study are to evaluate the disease control potential of Bacillus subtilis BS-99 strain against anthracnose disease causing by Colletotrichum gloeosporioides, on mango and papaya under field conditions. 3.Root disease control of export flowers by heat and effective microorganisms- In order to reduce crop diseases and reduction of chemical pesticide, the study in this project will conduct to use the steam with 70℃ and antagonists to control nematodes and root rot disease of export flowers. 4.Studies on soil amendments and antagonists for controlling Fusarium wilt of asparagus bean- Seeds and seedlings of asparagus bean will be treated with the potential biocontrol agents and planted into the pathogen-infested soils in the greenhouse and in the field. Based on the results obtained, we try to formulate a soil amendment with antagonists for controlling Fusarium wilt of asparagus bean in the field. 5. Application of probiotic Bacillus subtilis biofungicide in intergrated management of citrus canker disease- The completion of the project should provide a patentable proprietary technology for producing FNF from available B. subtilis biofungicides useful for sustainable agricultural production. 6.Development and application of broad-spectrum microbial agents for controlling of plant diseases- develop formulations of fluorescent Pseudomonas to control plant bacterial diseases. 7.Application of extracts of poisonous plant for control of nematode disease- Effective ingredients would be tested under greenhouse condition for its potential for future application. 8.Studies on the management of plant-parasitic nematode disease by using of cyanogenic bacteria. 9. 10. Developing sustainable IPM strategy for the control of legume pod borer, Maruca vitrata in vegetable legumes-(1) developing and standardizing reproducible mass culturing techniques for A. taragamae, (2) laboratory and greenhouse studies on the nature and extent of the parasitism by A. taragamae on LPB larvae viz., the most suitable host larval stage, effect of host larval population density on parasitism and effect of temperature, to optimize the conditions for field release, (3) testing the individual and combined effectiveness of N. maculosa and A. taragamae against LPB larvae field conditions, (4) testing the mating disruption and mass trapping potential of the LPB sex pheromones under controlled and open environment conditions using new pheromone formulations, (5) testing the effectiveness of MaviMNPV formulations alone and in combination with Bt and neem and (6) developing and testing the IPM with parasitoids, MaviMNPV, neem, Bt and sex pheromones in field experiments with yard-long bean. Thus, this proposal aimed to exploit the bio-control based IPM strategy to combat the LPB in vegetable legumes, which would not only reduce the insecticide use but also enhance the efficiency of natural enemies in field conditions. 11. Releases and evaluation of the effectiveness of a predtor, Cybocephalus binotatus, in controlling the cycad scale, Aulacaspis yasumatsui- In order to establish a natural enemy population as the biological control on the field, the mass propagation of C. japonicus will be conducted. In addition, the capture ability of C. japonicus to A. yasumatsui will be studied as the reference for mass release. The results will provide a completed pest manage model and reduce the total cost of controlling A. yasumatsui, benefiting the whole nation and avoiding the extinction of Cycad forest in Taitung.本計畫的目的在於： 1. 植物傷口癒合防護劑之研發與應用-篩選有益傷口癒合防護之微生物與拮抗菌,開發評估平台並合成傷口癒合防護劑。 2. 利用枯草桿菌生物製劑防治芒果及木瓜炭疽病可行性之探討-研究目標將以實驗室篩選出之本土優良拮抗枯草桿菌BS-99菌株，進行芒果及木瓜炭疽病的田間防治試驗。 3. 利用有益微生物與熱能防治外銷花卉根部病害之研究-利用熱能及本土性之微生物來減少洋秸梗病害及降低化學藥劑之使用。 4. 利用土壤添加物及拮抗微生物防治長豇豆萎凋病-以拮抗微生物配合土壤添加物，嘗試配合種子粉衣、剪胚軸接種或育苗介質混菌等方法，將拮抗微生物導入長豇豆幼苗，供作田間防治之用。 5. 益生性枯草桿菌在柑橘潰瘍病綜合防治管理之應用-完成預期可建立完成可專利化之「枯草桿菌生物殺菌劑泛用性FNF產製與應用」智財權技術，其推廣應用後除可強化我國農業生產之國際競爭力，並將加速永續農業政策推動之全面落實。 6. 植物病害防治用廣效性微生物製劑之研發與應用-發展螢光假單胞菌製劑來防治細菌性病害。 7. 利用有毒植物萃取液防治作物線蟲病害-評估有毒植物萃取液對不同植物寄生性線蟲之影響與應用於溫室之防治植物寄生性線蟲效果。 8. 利用產氰細菌防治作物線蟲病害之效果評估。 9. 芒果單爪葉蟎生物防治天敵效能評估-捕植蟎捕食芒果單爪葉蟎之功能反應之結果有助捕植蟎之田間利用，進而提供今後生物防治芒果單爪葉蟎之技術開發，落實非農藥防治及生物技術之利用，提高農友經濟收益，促進產品外銷及我國農產品競爭力。 10. 豆莢螟綜合防治技術之研究與開發(亞洲蔬菜中心-施瑞霖) (1) 開發豆莢螟寄生蜂Apanteles taragamae之大量飼養技術，及探討其生物學特性，包括最適寄生之寄主齡期、最適寄主密度、對溫度的敏感度等。 (2) 測試豆莢螟性費洛蒙之效果。 (3) 搭建可拆卸式網室，種植長豇豆，釋放豆莢螟及寄生蠅，並評估寄生蠅於網室長豇豆之寄生效果。 (4) 利用殺蟲效果較佳之豆莢螟核多角體病毒製劑BA與BH，單獨使用或與其他生物性藥劑(如蘇力菌、苦楝油)混用，評估豆莢螟核多角體於田間之防治效果。 (5) 綜合豆莢螟寄生蠅或寄生蜂、豆莢螟核多角體病毒、蘇力菌及性費洛蒙等防治技術，應用於長豇豆試驗田，並評估對豆莢螟之防治效果。 11. 釋放捕食性天敵防治蘇鐵白輪盾介殼蟲及其效果評估-了解生物天敵雙色出尾蟲捕食能力，做為釋放天敵數量之依據。大量飼養蘇鐵白輪盾介殼蟲之生物天敵雙色出尾蟲以建立蘇鐵白輪盾介殼蟲生物天敵族群，適合野外蘇鐵之生物防治模式，減少未來每年為防治蘇鐵白輪盾介殼蟲為害所付出之成本以完成蘇鐵白輪盾介殼蟲之綜合防治模式，使全國各地之蘇鐵白輪盾介殼蟲疫情得以控制，避免台東蘇鐵林滅絕

Investigate the Influence of the Quality of the Baking Strength to Camellia Oil

本研究以冷壓法壓榨油茶籽，並由不同烘焙溫度及時間調控茶油製程。溫度因子分為80、100、120、140及160°C五個水準，時間因子分為15、25及35分鐘三個水準。冷壓壓榨方式以兩段式間隔加壓進行，第一階段以加壓時間2.5秒停止12秒的方式，持續加壓至200kg/cm^2，並保持壓力狀態600秒。第二階段加壓方式相同，壓力由200kg/cm^2加壓至550kg/cm^2後，保持壓力狀態2200秒。壓榨所得茶油樣本經沉澱後，逐步進行各項理化特性分析。結果顯示，各試驗組之油樣品過氧化價都低於20毫當量（meq/kg），酸價都在2 mg KOH/g以內。褐變指數與油脂氧化安定性分析結果顯示，安定性最高的試驗組為160°C *35分鐘，所需時間最長可達到5.84 ± 0.17（hrs）。感官品評分析結果顯示，整體喜好度最高試驗組為140°C *15分鐘，其次為120°C *15分鐘。This study use cold pressing process to press camellia oil, and use different roasting temperature and time to control the process. The temperature factor is divided into five levels for 80, 100, 120, 140 and 160 °C. The time factor is divided into three levels for 15, 25, and 35 minutes. The cold pressing method use two-stage approach to provide the pressure intervals. The first stage use 2.5 seconds pressing time and 12 seconds stop time continuing increase the pressure up to 200 kg/cm^2 and maintain this pressure condition for 600 seconds. The second stage use the same pressurized way as the first stage. The pressure increased from 200 kg/cm^2 to 550 kg/cm^2 and maintains the pressure condition for 2200 seconds. After precipitation, the pressed Camellia oil sample had been analyzed for peroxine value, acid value, oil oxidation stability, sensory evaluation and browning index, and try to figure out the best process condition by the integrated analysis